Device with dynamic light scattering assembly

Abstract

An apparatus includes a processing chip and a dynamic light scattering component. The processing chip includes a fluid chamber and a light transmissive material adjacent the fluid chamber. The processing chip is removably positioned relative to the dynamic light scattering assembly. The dynamic light scattering assembly is used to direct light through the light transmissive material and into the fluid chamber. The dynamic light scattering assembly is also configured to receive light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber and thereby capture light scattering data. A processor determines the viscosity of the fluid in the fluid chamber based on the captured light scattering data. The processor also determines one or both of a particle size or a particle size distribution of particles in the fluid based on the captured light scattering data.

Description

[0001] Priority

[0002] This application claims priority to U.S. Patent Application No. 17 / 964,185, entitled "Apparatus with Dynamic Light Scattering Assembly," filed on October 12, 2022, which was published as U.S. Publication 2023 / 0035784 on February 2, 2023, the disclosure of which is hereby incorporated by reference in its entirety. Background Art

[0003] The subject matter discussed in this section should not be assumed to be prior art merely as a result of its mention in this section. Similarly, problems mentioned in this section or associated with the subject matter provided as background should not be assumed to have been previously recognized in the prior art. The subject matter in this section merely represents different approaches, which may themselves correspond to specific implementations of the claimed technology.

[0004] Some currently available techniques for manufacturing and formulating polynucleotide therapeutics (e.g., mRNA therapeutics, etc.) may expose the product to contamination and degradation. Some available centralized production may be too expensive, too slow, or vulnerable to contamination for use in therapeutic formulations that may include multiple polynucleotide species. Summary of the Invention

[0005] The development of scaled-up polynucleotide manufacturing, the production of single-patient doses, the elimination of contamination contact points, the input and process tracking for meeting clinical manufacturing requirements, and the use in point-of-care operations can advance the use of these therapeutic modalities. Microfluidic instruments and processes can provide advantages for achieving these goals. It may be desirable to measure the particle size and / or particle size distribution within a microfluidic system. Devices, systems, and methods for measuring the particle size and / or particle size distribution within a microfluidic system are described herein to overcome previously existing challenges and achieve the beneficial effects as described herein. Such microfluidic systems can be used to manufacture and formulate products containing biomolecules, such as therapeutic agents for personalized care.

[0006] It should be understood that all combinations of the foregoing concepts and additional concepts discussed in greater detail below (assuming such concepts are not mutually inconsistent) are considered to be part of the inventive subject matter disclosed herein and achieve the beneficial effects / advantages as described herein. Specifically, all combinations of the claimed subject matter that appear at the end of this disclosure are considered to be part of the inventive subject matter disclosed herein. Brief Description of the Drawings

[0007] Details of one or more specific embodiments are set forth in the accompanying drawings and the following detailed description. Other features, aspects, and advantages will become apparent from the detailed description, the drawings, and the claims, wherein:

[0008] Figure 1 A schematic diagram showing an example of a system including a microfluidic processing chip;

[0009] Figure 2 Shows Figure 1 An exploded perspective view of an example of the components of the system of;

[0010] Figure 3 Shows what can be incorporated into Figure 1 A top plan view of an example of a processing chip that can be incorporated into the system of;

[0011] Figure 4A Shows the Figure 3 Processing chip in a first operating state in a sectional side view;

[0012] Figure 4B Shows the Figure 3 Processing chip in a second operating state in a sectional side view;

[0013] Figure 4C Shows the Figure 3 Processing chip in a third operating state in a sectional side view;

[0014] Figure 4D Shows the Figure 3 Processing chip in a fourth operating state in a sectional side view;

[0015] Figure 4E Shows the Figure 3 Processing chip in a fifth operating state in a sectional side view;

[0016] Figure 4F Shows the Figure 3 Processing chip in a sixth operating state in a sectional side view;

[0017] Figure 5 Shows what can be incorporated into Figure 3 A perspective view of an example of a mixing stage that can be incorporated into the processing chip of;

[0018] Figure 6 A top plan view of an example of a portion of a processing chip incorporating a mixing stage;

[0019] Figure 7A Shows what can be incorporated into Figure 3 A schematic cross-sectional view of an example of a pressure sensing stage that can be incorporated into the processing chip of, where the elastic layer is in a non-deflected state;

[0020] Figure 7B Shows Figure 7A a schematic cross-sectional view of the pressure sensing stage, where the elastic layer is in a deflected state;

[0021] Figure 8 Shows Figure 7A a top plan view of a portion of the pressure sensing stage of

[0022] Figure 9 a perspective view of an exemplary assembly including a processing chip and a body for a dynamic light scattering stage;

[0023] Figure 10 Shows Figure 9 a top plan view of the assembly of

[0024] Figure 11 Shows Figure 9 a side elevation view of the assembly of

[0025] Figure 12 Shows Figure 9 an enlarged top plan view of the hybrid assembly in the processing chip of

[0026] Figure 13 Shows Figure 9 an enlarged plan view of the dynamic light scattering chamber in the processing chip of

[0027] Figure 14 Shows Figure 13 a perspective view of the dynamic light scattering chamber of

[0028] Figure 15 Shows Figure 13 another perspective view of the dynamic light scattering chamber of

[0029] Figure 16 Shows Figure 9 a perspective view of the body of the dynamic light scattering stage of the assembly, and a schematic representation of associated optical components;

[0030] Figure 17 Shows Figure 9 a perspective view of the body of the dynamic light scattering stage of the assembly, where the collimator and filter are separated from the body;

[0031] Figure 18 Shows Figure 9 a top plan view of the body of the dynamic light scattering stage of the assembly of

[0032] Figure 19 Shows Figure 9 a bottom plan view of the body of the dynamic light scattering stage of the assembly of

[0033] Figure 20 Shows along Figure 18Intercepted by line 20-20 Figure 9 Cross-sectional view of the main body of the dynamic light scattering stage of the component;

[0034] Figure 21 Shows the plot in Figure 9 Diagram showing an example of the autocorrelation function associated with the use of multimode optical fiber in the dynamic light scattering stage of the component;

[0035] Figure 22 Shows the plot in Figure 9 Diagram showing an example of the autocorrelation function associated with the use of single-mode optical fiber in the dynamic light scattering stage of the component;

[0036] Figure 23 Perspective view of a component including a processing chip and several main bodies for several corresponding dynamic light scattering stages;

[0037] Figure 24 Shows Figure 23 Top plan view of the component;

[0038] Figure 25 Perspective view of a measurement stage that can be incorporated into a processing chip;

[0039] Figure 26 Shows the representation of using Figure 25 Flowchart of an example of the process of measuring the viscosity of a processing chip using the measurement stage;

[0040] Figure 27 Shows the plot associated with Figure 25 Performed by the measurement stage of Figure 26 Diagram showing an example of the autocorrelation function of the process;

[0041] Figure 28 Shows the representation of using Figure 25 Another flowchart of the process of measuring the viscosity of a processing chip using the measurement stage;

[0042] Figure 29 Shows the plot of the gamma values that can be generated during the process performed by the measurement stage of Figure 25 Performed by the measurement stage of Figure 26 Diagram of an example;

[0043] Figure 30 Top plan view of another example of a processing chip;

[0044] Figure 31 Shows Figure 30 Another top plan view of the processing chip, with some features omitted for clarity;

[0045] Figure 32 Shows Figure 30An enlarged top-down plan view of a hybrid component of a processing chip; and

[0046] Figure 33 shows Figure 30 An enlarged top-down plan view of a measurement stage of a processing chip. DETAILED DESCRIPTION

[0047] In some aspects, devices and methods for processing therapeutic polynucleotides are disclosed herein. Specifically, these devices and methods can be closed-path devices and methods configured to minimize or eliminate manual operations during operation. The closed-path devices and methods can provide an almost completely sterile environment, and the components can provide a sterile path for the processing from an initial input (e.g., a template) to an output (e.g., a composite therapeutic agent). The materials input into the device (e.g., nucleotides and any chemical components) can be sterile; and can be input into the system with little to no manual interaction.

[0048] The devices and methods described herein can produce therapeutic agents with very fast cycle times and a very high degree of reproducibility. The devices described herein are configured to provide the synthesis, purification, dialysis, formulation, and concentration of one or more therapeutic compositions in a single integrated device. Alternatively, one or more of these processes can be performed in two or more devices as described herein. In some scenarios, the therapeutic composition comprises a therapeutic polynucleotide. Such therapeutic polynucleotides can include, for example, ribonucleic acid or deoxyribonucleic acid. The polynucleotide can include only natural nucleotide units or can include any kind of synthetic or semi-synthetic nucleotide units. All or some of the processing steps can be performed in an uninterrupted fluid processing path, which can be configured as one or a series of disposable microfluidic path devices, also referred to herein in some cases as processing chips or biochips (the chip does not necessarily have to be used for bio-related applications). This can allow for the synthesis (including formulation) of patient-specific therapeutic agents at the point of care (e.g., hospitals, clinics, pharmacies, etc.).

[0049] I. Overview of Systems Comprising Microfluidic Processing Chips

[0050] Figure 1An example of various components that can be incorporated into a system (100) is shown. The example system (100) includes a housing (103) that encloses a seating mount (115) capable of removably holding one or more microfluidic processing chips (111). In other words, the system (100) includes a component configured to removably accommodate the processing chip (111), where the processing chip (111) itself defines one or more microfluidic channels or fluid paths. Components of the system (100) that interact fluidically with the processing chip (111) (e.g., within the housing (103)) may include fluid channels or paths that need not be considered microfluidic (e.g., such fluid channels or paths are larger than the microfluidic channels or fluid paths in the processing chip (111)). In some forms, the processing chip (111) is provided and used as a disposable device, while the remainder of the system (100) is reusable. The housing (103) can be in the form of a chamber, a housing, etc. with an opening that can be closed (e.g., via a lid or a door, etc.) to thereby seal the interior. The housing (103) can enclose a thermal regulator and / or can be configured to be enclosed in a thermally regulated environment (e.g., a refrigeration unit, etc.). The housing (103) can form a sterile barrier. In some variations, the housing (103) can form a moist or humidity-controlled environment. Additionally or alternatively, the system (100) can be located in a cabinet (not shown). Such a cabinet can provide a temperature-regulated (e.g., refrigerated) environment. Such a cabinet can also provide air filtration and air flow management and can facilitate keeping reagents at a desired temperature throughout the manufacturing process. Additionally, such a cabinet can be equipped with UV lights for sterilizing the processing chip (111) and other components of the system (100). Various suitable features that can be incorporated into the cabinet housing the system (100) will be apparent to those skilled in the art in light of the teachings herein.

[0051] The seating mount (115) can be configured to secure the processing chip (111) using one or more pins or other components configured to hold the processing chip (111) in a fixed and predetermined orientation. The seating mount (115) can thus facilitate holding the processing chip (111) in the proper position and orientation relative to other components of the system (100). In the present example, the seating mount (115) is configured to hold the processing chip (111) in a horizontal orientation such that the processing chip (111) is parallel to the ground.

[0052] In some variations, the thermal control (113) can be positioned adjacent to the seating mount (115) to regulate the temperature of any processing chip (111) mounted in the seating mount (115). The thermal control (113) can include a thermoelectric component (e.g., a Peltier device, etc.) and / or one or more heat sinks for controlling the temperature of all or a portion of any processing chip (111) mounted in the seating mount (115). In some variations, more than one thermal control (113) can be included, such as to separately regulate the temperature of different regions in one or more areas of the processing chip (111). The thermal control (113) can include one or more thermal sensors (e.g., thermocouples, etc.) that can be used for feedback control of the processing chip (111) and / or the thermal control (113).

[0053] As Figure 1 shown, the fluid interface assembly (109) couples the processing chip (111) to a pressure source (117) to provide one or more paths for a fluid (e.g., a gas) at a positive or negative pressure to be conveyed from the pressure source (117) to one or more internal regions of the processing chip (111), as will be described in more detail below. Although only one pressure source (117) is shown, the system (100) can include two or more pressure sources (117). In some scenarios, the pressure can be generated by one or more sources other than the pressure source (117). For example, one or more vials or other fluid sources within the reagent storage rack (107) can be pressurized. Additionally or alternatively, reactions and / or other processes performed on the processing chip (111) can generate additional fluid pressure. In the present example, the fluid interface assembly (109) also couples the processing chip (111) to the reagent storage rack (107) to provide one or more paths for liquid reagents, etc. to be conveyed from the reagent storage rack (107) to one or more internal regions of the processing chip (111), as will be described in more detail below.

[0054] In some forms, pressurized fluid (e.g., gas) from at least one pressure source (117) reaches the fluid interface assembly (109) via the reagent rack (107), such that the reagent rack (107) includes one or more components interposed in the fluid path between the pressure source (117) and the fluid interface assembly (109). In some forms, one or more pressure sources (117) are directly coupled to the fluid interface assembly such that positive pressure fluid (e.g., positive pressure gas) or negative pressure fluid (e.g., suction or other negative pressure gas) bypasses the reagent rack (107) to reach the fluid interface assembly (109). Regardless of whether the fluid interface assembly (109) is interposed in the fluid path between the pressure source (117) and the fluid interface assembly (109), the fluid interface assembly (109) is capable of being removably coupled to the remainder of the system (100) such that at least a portion of the fluid interface assembly (109) can be removed for sterilization between uses. As described in more detail below, the pressure source (117) can selectively pressurize one or more chamber regions on the processing chip (111). Additionally or alternatively, the pressure source can also selectively pressurize one or more vials or other fluid storage containers held by the reagent rack (107).

[0055] The reagent rack (107) is configured to accommodate a plurality of fluid sample holders, each of the fluid sample holders being capable of holding a fluid vial or cassette configured to hold a reagent (e.g., nucleotide, solvent, water, etc.) for delivery to the processing chip (111). In some forms, one or more fluid vials, cassettes, or other storage containers in the reagent rack (107) can be configured to receive a product from the interior of the processing chip (111). Additionally or alternatively, a second processing chip (111) can receive a product from the interior of the first processing chip (111) such that one or more fluids are transferred from one processing chip (111) to another processing chip (111). In some such scenarios, the first processing chip (111) can perform a first dedicated function (e.g., synthesis, etc.), while the second processing chip (111) performs a second dedicated function (e.g., encapsulation, etc.). The reagent rack (107) of this example includes multiple pressure lines and / or a manifold configured to divide one or more pressure sources (117) into multiple pressure lines that can be applied to the processing chip (111). Such pressure lines can be controlled independently or jointly (in sub - combinations).

[0056] The fluid interface assembly (109) may include multiple fluid lines and / or pressure lines, where each such line includes a biased (e.g., spring-loaded) retainer or tip that individually and independently drives each fluid line and / or pressure line to the processing chip (111) when the processing chip (111) is held in the seating mount (115). Any associated tubing (e.g., fluid lines and / or pressure lines) may be part of the fluid interface assembly (109) and / or may be connected to the fluid interface assembly (109). In some forms, each fluid line includes a flexible tube connected between the reagent rack (107) and the processing chip (111), and the reagent rack couples the vial to the tube in a locking engagement (e.g., ferrule) via a connector. In some forms, the ends of the fluid line / pressure line may be configured to seal against the processing chip (111), for example, at corresponding sealed ports formed in the processing chip (111), as described below. In this example, the connection between the pressure source (117) and the processing chip (111) and the connection between the vial in the reagent rack (107) and the processing chip (111) both form sealed closed paths that are isolated when the processing chip (111) is seated in the seating mount (115). Such sealed closed paths may provide protection against contamination when processing therapeutic polynucleotides.

[0057] The vials in the reagent rack (107) may be pressurized (e.g., >1 atm pressure, such as 2 atm, 3 atm, 5 atm or higher). In some forms, the vials are pressurized by the pressure source (117). Thus, negative or positive pressure may be applied. For example, a fluid vial may be pressurized to about 1 psig to about 20 psig (e.g., 5 psig, 10 psig, etc.). Alternatively, a vacuum (e.g., about -7 psig or about 7 psig) may be applied at the end of the process to draw the fluid back into the vial (e.g., the vial serving as a reservoir). The fluid vial may be driven at a pressure lower than, for example, the pneumatic valve described below, which may prevent or reduce leakage. In some variations, the pressure difference between the fluid valve and the pneumatic valve may be between about 1 psi and about 25 psi (e.g., about 3 psi, about 5 psi, 7 psi, 10 psi, 12 psi, 15 psi, 20 psi, etc.).

[0058] The system (100) of this example further includes a magnetic field applicator (119) configured to form a magnetic field at the area of the processing chip (111). The magnetic field applicator (119) may include a movable head that is capable of operating to move the magnetic field so as to selectively isolate products that adhere to magnetic capture beads within the vials or other storage containers in the reagent rack (107).

[0059] The system (100) of this example also includes one or more sensors (105). In some forms, such sensors (105) include one or more cameras and / or other types of optical sensors. Such sensors (105) can sense one or more of barcodes, the liquid level within a fluid vial held within a reagent storage rack (107), fluid movement within a processing chip (111) mounted within a seating mount (115), and / or other optically detectable conditions. In forms where the sensor (105) is used to sense barcodes, such barcodes can be included on the vials of the reagent storage rack (107) such that the sensor (105) can be used to identify the vials within the reagent storage rack (107). In some forms, a single sensor (105) is positioned and configured to simultaneously view such barcodes on the vials in the reagent storage rack (107), the liquid level in the vials in the reagent storage rack (107), fluid movement within a processing chip (111) mounted within a seating mount (115), and / or other optically detectable conditions. In some other forms, more than one sensor (105) is used to view such conditions. In some such forms, different sensors (105) are positioned and configured to view corresponding optically detectable conditions separately such that the sensors (105) can be dedicated to specific corresponding optically detectable conditions.

[0060] In forms where the sensor (105) includes at least one optical sensor, visual / optical markers can be used to estimate the yield. For example, fluorescence can be used to detect process yield or residual material by labeling with fluorophores. Additionally or alternatively, dynamic light scattering (DLS) can be used to measure the particle size distribution within a portion of the processing chip (111) (e.g., such as a mixing portion of the processing chip (111)). In some variations, the sensor (105) can use one or two optical fibers to transmit light (e.g., a laser) to the processing chip (111) and detect the light signal emitted from the processing chip (111), thereby providing measurement results. In forms where the sensor (105) optically detects process yield or residual material, etc., the sensor (105) can be configured to detect visible light, fluorescence, ultraviolet (UV) absorption signals, infrared (IR) absorption signals, and / or any other suitable type of optical feedback.

[0061] In a form where the sensor (105) includes at least one optical sensor configured to capture video images, such a sensor (105) may record at least some activities on the processing chip (111). For example, the entire run for synthesizing and / or processing a material (e.g., therapeutic RNA) may be recorded by one or more video sensors (105), including a video sensor (105) capable of visualizing the processing chip (111) (e.g., from above). The processing on the processing chip (111) may be visually tracked, and the video recording may be retained for later quality control and / or processing. Thus, the video recording of the processing may be saved, stored, and / or transmitted for subsequent viewing and / or analysis. Additionally, as will be described in more detail below, the video may be used as a real-time feedback input that may affect the processing using at least visually observable conditions captured in the video.

[0062] The system (100) of this example is controlled by a controller (121). The controller (121) may include one or more processors, one or more memories, and various other electronic components, which will be apparent to those skilled in the art in reference to the teachings herein. In some forms, one or more components of the controller (121) (e.g., one or more processors, etc.) are embedded within the system (100) (e.g., housed within the housing (103)). Additionally or alternatively, one or more components of the controller (121) (e.g., one or more processors, etc.) may be detachably attached or detachably connected to other components of the system (100). Thus, at least a portion of the controller (121) may be removable. Additionally, in some forms, at least a portion of the controller (121) may be remote from the housing (103).

[0063] The control of the controller (121) may include activating the pressure source (117) to apply pressure through the processing chip (111) to drive fluid movement, and other tasks. The controller (121) may be entirely or partially external to the housing (103); or entirely or partially inside the housing (103). The controller (121) may be configured to receive user input via the user interface (123) of the system (100); and provide output to the user via the user interface (123). In some forms, the controller (121) is fully automated and does not require user input. In some such forms, the user interface (123) may only provide output to the user. The user interface (123) may include a monitor, a touch screen, a keyboard, and / or any other suitable features. The controller (121) may coordinate processing, including moving one or more fluids onto the processing chip (111), mixing one or more fluids on the processing chip (111), adding one or more components to the processing chip (111), metering the fluids on the processing chip (111), regulating the temperature of the processing chip (111), applying a magnetic field (e.g., when using magnetic beads), etc. The controller (121) may receive real-time feedback from the sensor (105) and execute a control algorithm based on such feedback from the sensor (105). Such feedback from the sensor (105) may include, but is not limited to, the identification of the reagent in the vial in the reagent storage rack (107), the detected liquid level in the vial in the reagent storage rack (107), the detected movement of the fluid on the processing chip (111), the fluorescence of the fluorophore in the fluid on the processing chip (111), etc. The controller (121) may include software, firmware, and / or hardware. The controller (121) may also communicate with a remote server, for example, to track the operation of the device, reorder materials (e.g., components such as nucleotides, processing chips (111), etc.), and / or download protocols, etc.

[0064] Figure 2 Examples of certain forms that various components of the system (100) may adopt are shown. Specifically, Figure 2 A reagent storage rack (150), a fluid interface assembly (152), a seating mount (154), a thermal control (156), and a processing chip (200) are shown. The reagent storage rack (150), the fluid interface assembly (152), the seating mount (154), the thermal control (156), and the processing chip (200) of this example may be constructed and operate respectively in the same way as the reagent storage rack (107), the fluid interface assembly (109), the seating mount (115), the thermal control (113), and the processing chip (111) described above. These components are fixed relative to the base (180). A set of rods (182) supports the reagent storage rack (150) on the fluid interface assembly (152).

[0065] As Figure 2As shown, a set of optical sensors (160) are positioned at four corresponding locations along a base (180). The optical sensors (160) can be configured and operated like the aforementioned sensors (105). The optical sensors (160) can include off-the-shelf cameras or any other suitable type of optical sensors. The optical sensors (160) are positioned such that fluid vials held within a reagent rack (150) are within the field of view of one or more of the optical sensors (160). Additionally, a processing chip (200) is within the field of view of one or more of the optical sensors (160). Each optical sensor (160) is movably fixed to the base (180) via a corresponding track (184) (e.g., in a gantry arrangement) such that each optical sensor (160) is configured to translate laterally along each corresponding track (184). A linear actuator (186) is fixed to each optical sensor (160) and is thereby capable of operating to drive the lateral translation of each optical sensor (160) along the corresponding track (184). Each actuator (186) can be in the form of a drive belt, a drive chain, a drive cable, or any other suitable type of structure. A controller (121) can drive the operation of the actuators (186). The optical sensors (160) can move along the tracks (184) during operation of the system (100) to facilitate viewing of appropriate regions of vials within the reagent rack (150) and / or the processing chip (200). In some scenarios, the optical sensors (160) move in unison along the corresponding tracks (184). In some other scenarios, the optical sensors (160) move independently along the corresponding tracks (184).

[0066] Although the optical sensors (160) are shown as mounted to the base (180) in Figure 2 , the optical sensors (160) can be positioned elsewhere within the system (100), as a supplement to or in place of being mounted to the base (180). For example, some forms of the reagent rack (107) can include one or more optical sensors (160) that are positioned and configured to provide a top field of view. In some such forms, such optical sensors (160) can be mounted to tracks, movable cantilevers, or other structures that allow such optical sensors (160) to be repositioned during operation of the system (100). Other suitable locations where the optical sensors (160) can be positioned will be apparent to those skilled in the art with reference to the teachings herein. Although not shown, the system (100) can also include one or more light sources (e.g., electroluminescent panels, etc.) to provide illumination that aids the optical sensing of the optical sensors (160).

[0067] In some configurations, one or more mirrors are used to facilitate visualization of components of the system (100) by the optical sensor (160). Such mirrors can allow the optical sensor (160) to view components of the system (100) that might otherwise be outside the field of view of the sensor (160). Such mirrors can be arranged to be directly adjacent to the optical sensor (160). Additionally or alternatively, such mirrors can be arranged adjacent to one or more components of the system (100) that are to be viewed by the optical sensor (160).

[0068] In use of the system (100), an operator can select a protocol to run (e.g., select from a library of preset protocols), or a user can input a new protocol (or modify an existing protocol) via the user interface (123). Depending on the protocol, the controller (121) can indicate which type of processing chip (111) to use, what the contents of the vials in the reagent rack (107) should be, and where to place the vials in the reagent rack (107). The operator can load the processing chip (111) into the seating mount (115); and load the required reagent vials and output vials into the reagent rack (107). The system (100) can confirm the presence of the required peripherals, identify the processing chip (111), and scan the identifiers (e.g., barcodes) of each reagent and product vial in the reagent rack (107) to facilitate vial matching to the reagent list for the selected protocol. After confirming the starting materials and equipment, the controller (121) can execute the protocol. During execution, valves and pumps are actuated to deliver reagents, the reagents are mixed, the temperature is controlled, and reactions occur, measurements are made, and then the product is pumped to a target vial in the reagent rack (107).

[0069] II. Examples of Processing Chips

[0070] Figure 3 and Figures 4A to 4F An example of the processing chip (200) is shown in more detail. In combination with the rest of the system (100), the processing chip (200) can be used to provide in vitro synthesis, purification, concentration, formulation, and analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides. As Figure 3As shown, the processing chip (200) of this example includes a plurality of fluid ports (220). Each fluid port (220) has an associated fluid channel (222) formed in the processing chip (200) such that fluid delivered to the fluid port (220) will flow through the corresponding fluid channel (222). As described in more detail below, each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from a fluid interface assembly (109). In this example, each fluid channel (222) leads to a valve chamber (224) that can be operated to selectively block or allow fluid from the corresponding fluid channel (222) to be further conveyed along the processing chip (200), as will be described in more detail below.

[0071] Also as Figure 3 shown, the processing chip (200) of this example includes a plurality of additional chambers (230, 250, 270) that can be used to serve different purposes during the production of a therapeutic composition as described herein. By way of example only, such additional chambers (230, 250, 270) can be used to provide synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions; or perform any other suitable function. Fluid can be conveyed from one chamber (230) to another via a fluid connector (232). In some configurations, the fluid connector (232) can operate between an open state and a closed state like a valve (e.g., similar to the valve chamber (224)). In some other configurations, the fluid connector (232) remains open throughout the process of preparing the therapeutic composition. In this example, the chamber (230) is used to provide the synthesis of polynucleotides, although the chamber (230) can alternatively be used for any other suitable purpose.

[0072] In Figure 3In the example shown, another valve chamber (234) is interposed between one of the chambers (230) and one of the chambers (250) such that fluid can be selectively transferred from the chamber (230) to the chamber (250). The chambers (250) are provided in pairs and are coupled to each other such that the processing chip (200) can transfer fluid back and forth between the chambers (250). Although a pair of chambers (250) are provided in this example, any other suitable number of chambers (250) can be used, including only one chamber (250) or more than two chambers (250). The chambers (250) can be used to provide purification of the fluid and / or can be used for any of the other various purposes described herein; and can have any suitable configuration. In the form where the chambers (250) are used for purification, the chambers (250) can include a material configured to absorb a selected portion from the fluid mixture in the chamber (250). In some such forms, the material can include a cellulose material that can selectively absorb double-stranded mRNA from the mixture. In some such forms, the cellulose material can be inserted into only one of a pair of chambers (270) such that after mixing the fluid from the first chamber (250) of the pair to the second chamber (250), the mRNA and / or some other components can be effectively removed from the fluid mixture, and the fluid mixture can then be transferred to another pair of chambers (250) further downstream for further processing or output. Alternatively, the chambers (250) can be used for any other suitable purpose.

[0073] An additional valve chamber (252) is interposed between each chamber (250) and the corresponding chamber (270) such that fluid can be selectively transferred from the chamber (250) to the chamber (270) via the valve chamber (252). The chambers (270) are also coupled to each other such that the processing chip (200) can transfer fluid back and forth between the chambers (270). The chambers (270) can be used to provide mixing of the fluid and / or can be used for any of the other various purposes described herein; and can have any suitable configuration.

[0074] As Figure 3 shown, the chambers (270) are also coupled to additional fluid ports (221) via corresponding fluid channels (223) and valve chambers (225). The fluid ports (221), fluid channels (223), and valve chambers (225) can be constructed and operated like the fluid ports (220), fluid channels (222), and valve chambers (224) described above. In some forms, the fluid ports (221) are used to transfer additional fluid to the chambers (270). Additionally or in an alternative, the fluid ports (221) can be used to transfer fluid from the processing chip (200) to another device. For example, fluid from the chambers (270) can be directly transferred via the fluid ports (221) to another processing chip (200), to one or more vials in the reagent storage rack (107), or elsewhere.

[0075] The processing chip (200) further includes a number of storage chambers (260). In this example, each storage chamber (260) is configured to receive and store fluids that are transferred to or from the corresponding chamber (250, 270). Each storage chamber (260) has a corresponding inlet valve chamber (262) and an outlet valve chamber (264). Each inlet valve chamber (262) is interposed between the storage chamber (260) and the corresponding chamber (250, 270) and can thus be operated to allow or prevent fluid flow between the storage chamber (260) and the corresponding chamber (250, 270). Each outlet valve chamber (264) can be operated to meter the fluid flow between the storage chamber (260) and the corresponding fluid port (266). In some forms, each fluid port (266) is configured to transfer fluid from a corresponding vial in the reagent storage rack (107) to the corresponding storage chamber (260). Additionally or alternatively, each fluid port (266) can be configured to transfer fluid from the corresponding storage chamber (260) to the corresponding vial in the reagent storage rack (107). In this example, the storage chambers (260) are used to provide metering of fluids transferred to and / or from the processing chip (200). Alternatively, the storage chambers (260) can be used for any other suitable purpose, including but not limited to pressurizing fluids transferred to and / or from the processing chip (200).

[0076] Also as Figure 3 shown, the processing chip (200) of this example includes a plurality of pressure ports (240). Each pressure port (240) has an associated pressure channel (244) formed in the processing chip (200) such that pressurized gas transferred through the pressure port (240) will be further transferred through the corresponding pressure channel (244). As described in more detail below, each pressure port (240) is configured to receive pressurized gas from a corresponding pressure line (208) from the fluid interface assembly (109). In this example, each pressure channel (244) leads to a corresponding chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) to provide valve actuation or peristaltic pumping through such chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270), as described in more detail below.

[0077] The processing chip (200) may further include electrical contacts, pins, pin sockets, capacitive coils, inductive coils, or other features configured to provide electrical communication with other components of the system (100). In Figure 3In the example shown, the processing chip (200) includes an electroactive region (212) having such electrocommunication characteristics. The electroactive region (212) may also include electronic circuits and other electronic components. In some forms, the electroactive region (212) may provide communication of power, data, etc. Although the electroactive region (212) is shown at a specific location on the processing chip, the electroactive region (212) may alternatively be located at any other suitable one or more locations. In some forms, the electroactive region (212) is omitted.

[0078] As Figures 4A to 4F shown, the processing chip (200) further includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306). As described in more detail below, some forms of the elastic layer (302) are in the form of a flexible film. The first plate (300) has an upper surface (210) and a lower surface (310), where the lower surface (310) faces the elastic layer (302). The second plate (304) has an upper surface (312) and a lower surface (314), where the upper surface (312) faces the elastic layer (302); and the lower surface (314) faces the third plate (306). The elastic layer (302) is thus interposed between the first plate (300) and the second plate (304). In this example, another elastic layer (316) is also interposed between the second plate (304) and the third plate (306), although the elastic layer (316) is optional.

[0079] The plates (300, 304, 306) of this example are substantially translucent to visible light and / or ultraviolet light. "Substantially translucent" means that at least 90% (including 100% in some cases) of the light passes through the material compared to a translucent material. In some variations, one or more of the plates (300, 304, 306) may include a material that is substantially transparent to visible light and / or ultraviolet light. "Substantially" translucent means that at least 90% (including 100% in some cases) of the light passes through the material compared to a completely transparent material. Also, for example, one or more of the plates (300, 304, 306) may provide transmission of ultraviolet light having a wavelength of about 260 nm with a transmittance in the range of about 0.2% to about 20%, including about 0.4% to about 15%, or including about 0.5% to about 10%.

[0080] The plates (300, 304, 306) of this example are also rigid. In some other forms, one or more of the plates (300, 304, 306) are semi-rigid. The plates (300, 304, 306) can include glass, plastic, silicone, and / or any other suitable material. In some forms, one or more of the plates (300, 304, 306) are formed as a laminate of two or more layers of material, such that each plate (300, 304, 306) does not necessarily have to be formed as a single homogeneous continuous material. The material forming one of the plates (300, 304, 306) can also be different from the material forming the other plates (300, 304, 306).

[0081] The elastomeric layer (302) of this example is formed as a liquid-impermeable flexible membrane. In some forms, the elastomeric layer (302) is gas-permeable, although liquid-impermeable. In some forms of this type, certain regions of the elastomeric layer (302) are treated to be gas-permeable, while the untreated regions of the elastomeric layer (302) are gas-impermeable. As described below, the elastomeric layer (302) can be used to drive fluid through the processing chip (200) via a peristaltic pumping action. Also as described below, the elastomeric layer (302) can be used to provide valves at various positions along the processing chip (200). In some forms, a single piece of elastomeric material spans the width of the processing chip (200) to form the elastomeric layer (302). In some other forms, two or more discrete pieces of elastomeric material are used to form the elastomeric layer (302), such discrete pieces of elastomeric material being positioned at different locations across the width of the processing chip (200). By way of example only, the elastomeric layer (302) can include a membrane comprising a polydimethylsiloxane (PDMS) elastomeric membrane.

[0082] As Figures 4A to 4F Best seen in, the first plate (300) and the second plate (304) cooperate to define a plurality of chambers (320, 322, 324, 326), wherein the elastomeric layer (302) divides each chamber (320, 322, 324, 326) into a corresponding upper chamber region (330) and a lower chamber region (332). Figure 3 The chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) shown can be constructed and operated like Figures 4A to 4F the chambers (320, 322, 324, 326) shown. For example, chamber (320) can be similar to chamber (264), chamber (322) can be similar to chamber (260), chamber (324) can be similar to chamber (262), and chamber (326) can be similar to chamber (250).

[0083] As Figures 4A to 4FAs shown, a fluid port (220) is formed through the first plate (220). A corresponding opening (342) is formed through a region of the elastomeric layer (302) that is located below the fluid port (220). A fluid channel (222) extends from the opening (342) to a lower chamber region (332) of the first chamber (320). As described above, the fluid port (220) is configured to receive a fluid line (206) from a fluid interface assembly (109). The distal end of the fluid line (206) is configured to seal against a region of the elastomeric layer (302) that is exposed by the fluid port (220) and to convey fluid (207) through the opening (342). In some configurations, a spring or other elastic member provides an elastic bias to the fluid line (206), pushing the distal end of the fluid line (206) against the region of the elastomeric layer (302) that is exposed by the fluid port (220) to maintain the seal. Fluid (207) from the fluid line (206) reaches the lower chamber region (332) of the first chamber (320) via the fluid channel (222). As described in more detail below, this fluid (207) can be further conveyed from the first chamber (320) to other chambers (322, 324, 326) through a peristaltic pumping action provided by the elastomeric layer (302). After reaching the fourth chamber (326), the fluid (207) can be further conveyed to other chambers or other features in the processing chip (200), can be conveyed to storage vials in a reagent rack (107), or can be otherwise processed. The path for the fluid (207) thus need not terminate at the fourth chamber (326). It should also be understood that Figure 3 any other fluid ports (221, 266) shown can be configured and operated like Figures 4A to 4F the fluid port (220) shown.

[0084] The pressure port (240) is formed through the first plate (220). A corresponding opening (344) is formed through a region of the elastic layer (302) that is located below the fluid port (240). The pressure channel (244) extends from the opening (344) to the upper chamber region (330) of the first chamber (320). As described above, the pressure port (240) is configured to receive a pressure line (208) from the fluid interface assembly (109) so as to receive pressurized gas from a pressure source (117). The distal end of the pressure line (208) is configured to seal against a region of the elastic layer (302) that is exposed by the pressure port (240) and to transmit positive or negative pressure gas through the opening (344). In some configurations, a spring or other elastic member provides an elastic bias to the pressure line (208), pushing the distal end of the pressure line (208) against the region of the elastic layer (302) that is exposed by the pressure port (240) so as to maintain the seal. The positive or negative pressure gas from the pressure line (208) reaches the upper chamber region (330) of the fourth chamber (326) via the pressure channel (244).

[0085] Although Figures 4A to 4F only one pressure line (208) is shown as being coupled to the processing chip (200), the processing chip (200) may have a plurality of coupled pressure lines (208), where such pressure lines (208) independently apply positive or negative pressure to corresponding chambers (320, 322, 324, 326) of the processing chip (200). In some configurations, one or more of the chambers (320, 322, 324, 326) have their own dedicated pressure line (208) and corresponding pressure channel (244). Additionally or alternatively, one or more of the chambers (320, 322, 324, 326) may share a common pressure line (208) via the same pressure channel (244) or via separate pressure channels (244). Although Figures 4A to 4F the pressure channel (244) is shown as being formed through the second plate (304), some pressure channels (244) (or regions of the pressure channel (244)) may be formed by the first plate (300). For example, some pressure channels 244 (or regions of the pressure channel 244) may be formed between a recess in the lower surface of the first plate (300) and the upper surface of the elastic layer (302).

[0086] A. Examples of Valve Modulation and Peristaltic Pumping Driven via an Elastomeric Layer

[0087] As described above, the elastic layer (302) can be operated to drive fluid through the processing chip (200) by a peristaltic pumping action; and to block the movement of fluid through the processing chip (200) by providing a valve-like action. Examples of such operations are provided in Figures 4A to 4FThe sequences shown. In this example, chambers (320, 324) are used as valve chambers, while chamber (322) is used as a metering chamber. Chamber (326) is used as a working chamber such that synthesis, purification, dialysis, compounding, concentration, or some other process is performed in chamber (326). This construction, arrangement, and use of chambers (320, 322, 324, 326) are provided as an illustrative example. Chambers (320, 322, 324, 326) may alternatively be constructed, arranged, and used in other ways.

[0088] Figure 4A It is shown that the processing chip (200) is in a state where fluid has not been delivered to the processing chip (200) and pressurized gas has not been delivered to the processing chip (200). In Figure 4B , positive pressure gas is delivered to the upper chamber region (330) of chamber (320), negative pressure gas is delivered to the upper regions (330) of chambers (324, 322), and fluid (207) is delivered to chambers (320, 322). In this state, the positive pressure gas deforms a portion of the elastic layer (302) in chamber (324) such that the elastic layer (302) is seated against the surface of the lower chamber region (332) of chamber (324). This seating of the elastic layer (302) against the surface of the lower chamber region (332) of chamber (324) prevents fluid (207) from entering chamber (324), such that chamber (324) operates like a closed valve in the Figure 4B shown state. The negative pressure gas in the upper chamber regions (330) of chambers (320, 322) causes the corresponding portions of the elastic layer (302) in chambers (320, 322) to deform and be seated against the upper chamber regions (330) of chambers (320, 322). This allows fluid (207) to occupy the entire capacity of chambers (320, 322).

[0089] After reaching the Figure 4B shown state, positive pressure gas is delivered to the upper chamber region (330) of chamber (322), while the pneumatic states of chambers (320, 324) may remain unchanged. This results in the Figure 4C shown state. As shown, the positive pressure gas deforms a portion of the elastic layer (302) in chamber (320) such that the elastic layer (302) is seated against the surface of the lower chamber region (332) of chamber (320). This seating of the elastic layer (302) against the surface of the lower chamber region (332) of chamber (320) drives fluid (207) out of chamber (320) and causes chamber (320) to operate like a closed valve in the Figure 4C shown state. However, the volume of fluid (207) in chamber (322) in the Figure 4Cis not affected in the shown state. Chamber (322) can thus be used to provide metering of fluid (207) such that only an exact predetermined volume of fluid (207) is further conveyed along the processing chip (200). By way of example only, such metered volumes can be of the order of about 10 nL, 20 nL, 25 nL, 50 nL, 75 nL, 100 nL, 1 microliter, 5 microliters, etc.

[0090] Once the appropriate metered volume is achieved, negative pressure gas is conveyed to the upper chamber region (330) of chambers (320, 322), while the pneumatic state of chambers (324, 326) can remain unchanged. This results in Figure 4D the shown state. As shown, the negative pressure gas in the upper chamber region (330) of chambers (324, 326) causes the corresponding portions of the elastic layer (302) in chambers (324, 326) to deform and rest against the surface of the upper chamber region (330) of chambers (324, 326). This effectively opens the valve formed by chamber (324) and places chamber (326) in a state to receive fluid (207). This also creates a negative pressure in chamber (324) that sucks fluid (207) from chamber (322) into chamber (324).

[0091] With the valve formed by chamber (324) in an open state, positive pressure gas is conveyed to the upper chamber region (330) of chamber (320), while the pneumatic state of chambers (322, 324, 326) can remain unchanged. This results in Figure 4E the shown state. As shown, the positive pressure gas in the upper chamber region (330) of chamber (322) causes the corresponding portions of the elastic layer (302) in chamber (322) to deform and rest against the surface of the lower chamber region (332) of chamber (322). This deformation of the elastic layer (302) drives fluid (207) out of chamber (322). Since the valve formed by chamber (320) is in a closed state and the valve formed by chamber (324) is in an open state, fluid (207) travels from chamber (322) into chamber (324). In this example, the capacity of chamber (322) is greater than the capacity of chamber (324) such that fluid (207) from chamber (326) overflows from chamber (322) into chamber (324).

[0092] Once fluid (207) has been conveyed from chamber (322) to chambers (324, 326), positive pressure gas is conveyed to the upper chamber region (330) of chamber (320), while the pneumatic state of chambers (324, 322, 326) can remain unchanged. This results in Figure 4FThe state shown. As shown, the positive pressure gas in the upper chamber region (330) of the chamber (324) causes the corresponding portion of the elastic layer (302) in the chamber (324) to deform and rest against the surface of the lower chamber region (332) of the chamber (324). This deformation of the elastic layer (302) drives the fluid (207) out of the chamber (324). Since the deformed portion of the elastic layer (302) in the chamber (324) effectively seals the chamber (324) from the chamber (324) (e.g., such that the chamber (324) operates like a valve in a closed state), the fluid (207) travels from the chamber (324) into the chamber (326).

[0093] At Figure 4F the stage shown, the fluid (207) has been discharged from the chambers (320, 332, 324), and the chamber (326) contains a precisely metered volume of the fluid (207) within the chamber (322). The fluid (207) in the chamber (326) can be further processed within the chamber (326) according to the teachings herein. Additionally or alternatively, the fluid (207) in the chamber (326) can be transferred to one or more other chambers in the processing chip (200), can be transferred to vials in the reagent rack (107), or can be otherwise processed. Regardless of what operation is performed on the fluid (207) after the fluid (207) has reached the chamber (326), it should be understood that the fluid (207) is conveyed sequentially along the chambers (320, 322, 324) to reach the chamber (326) via a peristaltic action generated by the elastic layer (302) in response to positive or negative pressure gas being conveyed to the upper chamber region (330) of the chambers (320, 322, 324, 326) in a specific sequence. Such peristaltic pumping may have particular advantages for moving fluids that may be viscous or contain suspended particles such as purification or capture beads. Such peristaltic pumping achieved through selective deformation of the elastic layer (302) may also be referred to as pneumatic barrier deflection or "pneumatic deflection".

[0094] In some scenarios, it may be desirable to remove air or other gases from one or more fluid paths in a processing chip (200). To achieve this, the processing chip (200) can include one or more chambers configured to provide fluid access or otherwise vent gases from the fluid paths. For example, such venting or evacuation can be performed as part of a perfusion process as fluid is initially introduced into the processing chip (200). Additionally or alternatively, such venting or evacuation can be performed to release gases generated in the fluid during the formation of a therapeutic composition. Such a venting or gas release chamber can be referred to as a "vacuum lid". In some configurations, at least the region of the elastomeric layer (302) located within the vacuum lid (if not the entire elastomeric layer (302)) is gas permeable (while still being liquid impermeable). A negative pressure gas can be applied to the upper chamber region (330) of the chamber that serves as the vacuum lid, and this negative pressure gas can draw air or gas from the fluid path through the corresponding region of the elastomeric layer (302). In some configurations, the upper chamber region (330) of the chamber that serves as the vacuum lid includes one or more protrusions or support features that prevent the corresponding region of the elastomeric layer (302) from fully seating against the surface of the upper chamber region (330) of the chamber that serves as the vacuum lid. This can further facilitate the evacuation of air or other gases via the vacuum lid.

[0095] B. Examples of Mixing Stages

[0096] Although the chamber (270) can be used to perform mixing of fluids (e.g., by repeatedly shuttling the fluid back and forth between chambers (270)), it may be desirable to provide differently configured mixing stages along the fluid paths leading to the chambers. Figure 5 An example of such a mixing stage (400) that can be incorporated into a processing chip (111, 200) is shown. The mixing stage (400) can also be referred to as a "mixer", such that the terms "mixer" and "mixing stage" should be understood interchangeably. The illustrated mixing stage (400) includes two fluid inlet channels (402, 404) that are offset from each other and are configured to convey one or more substances (e.g., biomolecular products, buffers, carriers, auxiliary components) that can be combined together. Although two inlet channels (402, 404) are shown, three or more (4, 5, 6, etc.) can be used and can converge within the same mixing stage (400). The fluid mixture can pass through the inlet channels (402, 404) under positive pressure. This pressure can be constant, variable, increasing, decreasing, and / or pulsating. The inlet channels (402, 404) can receive fluid from any of the various chambers or fluid ports described herein.

[0097] The inlet channels (402, 404) converge at an intersection point (406) leading to the merging channel (408). In this example, the merging channel (408) has a cross-sectional area that is smaller than the cross-sectional area of each inlet channel (402, 404). The reduced cross-sectional area can include a channel height that is less than the channel height of the inlet channels (402, 404) and / or a channel width that is less than the channel width of the inlet channels (402, 404). This reduced cross-sectional area can facilitate the mixing of the fluids introduced via the inlet channels (402, 404).

[0098] The first vortex mixing chamber (414) is positioned downstream of the merging channel (408), where fluid flows into the first vortex mixing chamber (414) via an inlet opening (410). The inlet opening (410) is positioned near one corner of the first vortex mixing chamber (414). An outlet opening (412) is positioned near the other corner of the first vortex mixing chamber (414). The first vortex mixing chamber (414) has a height and width that are greater than the height and width of the merging channel (408). These larger dimensions and the relative positioning of the inlet opening (410) and the outlet opening (412) can facilitate the formation of vortices within the first vortex mixing chamber (414). As the fluid flows through the first vortex mixing chamber (414), such vortices can further facilitate the mixing of the fluid.

[0099] The connecting channel (416) connects the first vortex mixing chamber (414) to the second vortex mixing channel (420). The connecting channel (416) has a height and width that are less than the height and width of the first vortex mixing chamber (414). The second vortex mixing channel (420) has a height and width that are greater than the height and width of the connecting channel (416). Fluid flows from the connecting channel (416) into the second vortex mixing channel (420) via an inlet opening (418) positioned near a corner of the second vortex mixing chamber (420). The fluid flows out of the second vortex mixing chamber (420) via an outlet opening (422) positioned at the other corner of the second vortex mixing chamber (420). The outlet opening (420) leads to the outlet channel (424). The outlet channel (424) has a height and width that are less than the height and width of the second vortex mixing chamber (420). The larger dimensions of the second vortex mixing chamber (420) (relative to the dimensions of the channels (416, 424)) and the relative positioning of the inlet opening (418) and the outlet opening (422) can facilitate the formation of vortices within the second vortex mixing chamber (420). As the fluid flows through the second vortex mixing chamber (420), such vortices can further facilitate the mixing of the fluid.

[0100] When the fluid exits the outlet channel (424), the fluid can be thoroughly mixed by the mixing stage (400). Such mixed fluid can be further conveyed to other chambers or ports for further processing. Although the mixing stage (400) of this example has two vortex mixing chambers (414, 420), other configurations may have only one vortex mixing chamber or more than two vortex mixing chambers.

[0101] Figure 6 An example of a region of a processing chip (500) incorporating two mixing stages is shown. In this example, a first fluid passes through a first inlet valve (510), then through a first restrictor (520) in the form of a serpentine channel, then through a first vacuum cover (530), and then reaches a first inlet (540) of the first mixing stage. A second fluid passes through a second inlet valve (512), then through a second restrictor (522) in the form of a serpentine channel, then through a second vacuum cover (532), and then reaches a second inlet (542) of the first mixing stage. The inlets (540, 542) converge to provide a single flow path through a merging channel (544) that leads to a first set (550) of vortex mixing chambers. The first set (550) of vortex mixing chambers can be constructed and operated like the aforementioned vortex mixing chambers (414, 420). Although four vortex mixing chambers are included in the first set (550) in this example, the first set (550) can alternatively have any other suitable number of vortex mixing chambers.

[0102] After flowing through the first set (550) of vortex mixing chambers, the fluid reaches a first inlet (560) of the second mixing stage. A third fluid passes through a third inlet valve (514), then through a third restrictor (524) in the form of a serpentine channel, then through a third vacuum cover (534), and then reaches a second inlet (562) of the second mixing stage. The inlets (560, 562) converge to provide a single flow path through a merging channel (564) that leads to a second set (552) of vortex mixing chambers. The second set (552) of vortex mixing chambers can be constructed and operated like the aforementioned vortex mixing chambers (414, 420). Although two vortex mixing chambers are included in the second set (552) in this example, the second set (552) can alternatively have any other suitable number of vortex mixing chambers.

[0103] After flowing through the second set (552) of vortex mixing chambers, the fluid passes through a fourth vacuum cover (536). After passing through the fourth vacuum cover (536), the fluid can be thoroughly mixed by the two sets (550, 552) of vortex mixing chambers; and any bubbles may have been removed by the vacuum covers (530, 532, 534, 536). The mixed fluid can be further conveyed to other chambers or ports for further processing after passing through the fourth vacuum cover (536).

[0104] In one example of how a processing chip (500) may be used, a polynucleotide (e.g., mRNA in water) may be introduced via a first inlet valve (510), while one or more delivery carrier molecules in a fluid medium (e.g., ethanol or some other fluid medium) may be introduced via a second inlet valve (512). These fluids may be mixed in a first set (550) of vortex mixing chambers to form complex nanoparticles. A diluent (e.g., a citrate-based buffer solution or other type of buffer) may be introduced via a third inlet valve (514) to provide pH adjustment as the diluent is mixed with the complex nanoparticles in a second set (552) of vortex mixing chambers. Other suitable ways of using the processing chip (500) will be apparent to those skilled in the art in light of the teachings herein.

[0105] The foregoing structures are examples of how mixing of fluids from different sources may be performed in a processing chip (111, 200, 500). It is contemplated that various other types of structures may be used to provide mixing of fluids from different sources in a processing chip (111, 200, 500).

[0106] C. Examples of Pressure Sensing Stages

[0107] In some scenarios, it may be desirable to provide one or more sensors capable of operating to sense fluid pressure in a processing chip (111, 200, 500). Figures 7A to 7B An example of a pressure sensing stage (700) including a portion of a processing chip (710), a camera (702), and a controller (121) is shown. In addition to including the features and functions described below, the processing chip (710) may also include any other features and functions described above in the context of the processing chips (111, 200, 500). In other words, the following teachings related to the pressure sensing stage (700) may be readily applied to any of the various processing chips (111, 200, 500) described herein.

[0108] The camera (702) of this example is positioned to provide a field of view (704) in which the camera (702) can capture an image of the optical features (760) of the processing chip (700). The controller (121) receives image signals from the camera (702) and processes those image signals to determine fluid pressure values, as described in more detail below. The controller (121) can also use at least such determined fluid pressure values to perform various algorithms, as will also be described in more detail below. In this example, the controller (121) of the pressure sensing stage (700) is the same controller (121) that is used to perform other operations in the system (100) as described above. In some other configurations, a separate controller is used to determine fluid pressure values based at least on image signals from the camera (702). In such a configuration, the separate controller can transmit those determined fluid pressure values to the controller (121) to perform pressure-based algorithms. Alternatively, the determined fluid pressure values can be utilized by any other suitable hardware component in any other suitable manner.

[0109] The processing chip (710) of this example includes a first plate (720), an elastomeric layer (730), a second plate (740), and a third plate (750). The elastomeric layer (730) is interposed between two plates (720, 740). The third plate (750) mates with the second plate (740) to define a channel (742) through which fluid can flow. The region of the channel (742) located Figures 7A to 7B on the left hand side of can be regarded as the fluid inlet port of the pressure sensing stage (700); while the region of the channel (742) located Figures 7A to 7B on the right hand side of can be regarded as the fluid outlet port of the pressure sensing stage (700). The plates (720, 740, 750) of the processing chip (710) can be constructed and operate like the plates (300, 304, 306) of the processing chip (200). Similarly, the elastomeric layer (730) of the processing chip (710) can be constructed and operate like the elastomeric layer (302) of the processing chip (200). Thus, the elastomeric layer (730) can extend across all or most of the width of the processing chip (710) such that the elastomeric layer (730) can also perform functions (e.g., valve modulation, peristaltic pumping, venting, etc.) in other chambers of the processing chip (710).

[0110] The second plate (740) defines an opening (744) that is fluidly coupled to the channel (742), such that the opening (744) exposes a portion (732) of the elastic layer (730) to the fluid in the channel (742). The first plate (720) defines an opening (722) that is aligned with the opening (744) of the second plate (740). In this example, since the opening (744) provides a path for the fluid in the channel (742) to reach the portion (732) of the elastic layer (730), and since the opening (722) provides a gap for the elastic layer (730) to deform, the portion (732) of the elastic layer (730) can achieve the deformation state as shown in Figure 7B when pressurized positively by the fluid within the channel (742).

[0111] The optical feature (760) is positioned on the top portion (732) of the elastic layer (730). The optical feature (760) is configured to deform together with the elastic layer (730). For example, as shown by the transition from Figure 7A (non-pressurized state) to Figure 7B (pressurized state), the elastic layer (730) and the optical feature (760) deform upward together along the central axis (CA) in response to the positive pressurization of the fluid within the channel (742). In this example, the central axis (CA) is perpendicular to the plane defined by the elastic layer (730) when the elastic layer (730) is in the non-deformed state ( Figure 7A ); and is positioned at the radial center of the opening (722). Figure 7B The pressurized state shown in can occur during any of the various operations described herein during the peristaltic driving of the fluid from one position upstream of the channel (742) to another position downstream of the channel (742). Additionally or alternatively, Figure 7B the pressurized state shown in can occur in various other scenarios, including but not limited to the fluid in the channel (742) coming from a pressurized fluid source in the reagent storage rack (107), ambient pressure changes, pressure losses due to piping, and / or various other conditions. In the case where the optical feature (760) is directly or indirectly within the field of view (704) of the camera (702), the camera (702) is operable to capture an image of the deformation of the optical feature (760) and transmit the image data to the controller (121). The controller (121) is operable to convert the image data into a pressure value indicative of the fluid pressure in the channel (742), as will be described in more detail below.

[0112] As the elastic layer (730) and the optical feature (760) deform together along the central axis (CA) in response to the positive pressurization of the fluid within the channel (742) ( Figure 7B), the elastic layer (730) and the optical features (760) may also deform along a lateral dimension (LD) that is transverse to the central axis (CA). As described in more detail below, the camera (702) and the controller (121) may be operable to track this "lateral deformation" specifically along the lateral dimension (LD) to determine the pressure of the fluid in the channel (742). Such lateral deformation of the elastic layer (730) and the optical features (760) may be tracked in addition to or in lieu of tracking deformation of the elastic layer (730) and the optical features (760) along the central axis (CA).

[0113] Although Figure 7B The elastic layer (730) and optical features (760) are shown deforming upward along the central axis (CA) in response to positive pressurization of the fluid in the channel (742), but there may also be scenarios where the elastic layer (730) and optical features (760) deform downward along the central axis (CA) in response to negative pressurization of the fluid in the channel (742). In such scenarios, the elastic layer (730) and optical features (760) may also achieve lateral deformation as described above. The camera (702) and controller (121) may therefore be operated to track this lateral deformation to determine the pressure of the fluid in the channel (742), regardless of whether the pressure is positive (resulting in deformation upward along the central axis (CA)) or negative (resulting in deformation downward along the central axis (CA)).

[0114] like Figure 8 As shown, the optical feature (760) spans the entire radial distance (D 1 Alternatively, the optical feature (760) may only span the entire radial distance (D 1 ). In some versions, it may be desirable to track lateral deformation of the elastic layer (730) via an annular region (762) of the optical feature (760). In other words, it may be desirable to track deformation of an annular region of the elastic layer (730) by optically tracking the optical feature (760) within the annular region (762) of the optical feature (760). In this example, the annular region (762) is offset radially outward from the central axis (CA); and radially inward from the outer periphery of the opening (722). The annular region (762) is defined within a first portion of the radial distance (D 2 ) and the radial distance (D 3 ). The annular region (762) is therefore located between these portions of the radial distance (D 2 , D 3 ) has a radial dimension (D 4 ).

[0115] By way of example only, the opening (722) can have an overall radial distance (D) in the range of from about 0.75 mm to about 3.5 mm 1 ). Also by way of example only, the range of the first partial radial distance (D 2 ) can be from about 0.2 mm to about 2.0 mm. Also by way of example only, the range of the second partial radial distance (D 3 ) can be from about 1.0 mm to about 3.0 mm. Also by way of example only, the radial dimension (D 4 ) of the annular region (762) can be from about 0.5 mm to about 2.25 mm. As another example, the optical feature (760) can take the form of concentric rings spaced from each other by a distance in the range of from about 50 microns to about 150 microns.

[0116] In this example, the optical feature (760) does not affect the elasticity of the elastic layer (730). In some forms, the optical feature (760) is adhered to the elastic layer (730) via an adhesive. In some other forms, the optical feature (760) is in the form of a film applied to the elastic layer (730). In some other forms, the optical feature (760) is directly printed on the elastic layer (730). In some other forms, the optical feature (760) is engraved on the elastic layer (730). In some other forms, the optical feature (760) is formed as a texture on the elastic layer (730). Alternatively, the optical feature (760) can be fixed to the elastic layer (730) or otherwise incorporated into the elastic layer in any other suitable manner. In some forms, the optical feature (760) spans the entire area of the portion (732) of the elastic layer (730) as defined by the overall radial distance (D 1 ) of the opening (722). In some other forms, the optical feature (760) is located only on one or more discrete regions of the portion (732) of the elastic layer (730) within the opening (722) and does not span the entire area of the portion (732) of the elastic layer (730) within the opening (722). For example, in some forms, the optical feature (760) is located only in Figure 8 the annular region (762) shown such that the optical feature (760) does not extend through the first partial radial distance (D 2 ) or through the space between the second partial radial distance (D 3 ) and the overall radial distance (D 1 ).

[0117] In this example, the pressure sensing portion (732) and the optical feature portion (760) of the elastic layer (730) are exposed to the ambient atmosphere such that the deformation of the elastic layer (730) and the optical feature portion (760) utilizes at least the difference between the fluid pressure in the channel (742) and the atmospheric pressure. In some other configurations, the region above the pressure sensing portion (732) and the optical feature portion (760) of the elastic layer (730) of the processing chip (700) can be enclosed and exposed to a fluid path pressurized by the system (100) at a known pressure level. In such scenarios, the controller (121) can measure the pressure of the fluid in the channel (742) relative to the known system-generated pressure level. This type of configuration can prevent changes in atmospheric pressure from affecting the pressure sensing process in the manner that might otherwise occur in configurations where the pressure sensing portion (732) and the optical feature portion (760) of the elastic layer (730) are exposed to the ambient atmosphere.

[0118] III. Examples of Particle Size Sensing via Dynamic Light Scattering

[0119] As described above, one or more processing chips (111, 200, 500) can be used to prepare polynucleotide therapeutics (e.g., mRNA therapeutics, etc.). For example, a polynucleotide such as mRNA in water can be mixed with one or more delivery carrier molecules in ethanol to form complex nanoparticles. In some scenarios where a processing chip (111, 200, 500) is used to prepare an mRNA therapeutic composition, it may be desirable to encapsulate the mRNA in particles having a diameter of about 100 nm. The encapsulation process can include adjusting the mixing of the mRNA and the delivery carrier molecules via a mixing stage such as the mixing stage (400) described above. Such delivery carrier molecules can include lipid-based molecules such as amino lipidated peptoids. During this process, the temperature of the mixing stage (400) can be controlled to a temperature or temperature range (e.g., between about 2 degrees Celsius and about 20 degrees Celsius) that is calibrated to enhance the mixing action for mixing in the mixing stage (400). The elevated mixing temperature can be based on the formulation being mixed (in some examples, the sequence of the mRNA and / or the delivery carrier) within the specific geometry of the mixing chamber.

[0120] As used herein, a "delivery carrier" refers to any substance that at least partially facilitates the in vivo, in vitro, or ex vivo delivery of a polynucleotide to a target cell or tissue (e.g., a tumor, etc.). Referring to something as a delivery carrier does not exclude the possibility that the delivery carrier also has a therapeutic effect. Some types of delivery carriers can provide additional therapeutic effects. In some configurations, the delivery carrier can be an amino lipidated peptoid delivery carrier that can at least partially encapsulate the mRNA.

[0121] Variations in the shape, flow rate, chemical composition, and mixing ratio of the mixing chambers (414, 420, 550, 552) can all affect the particle size and / or particle size distribution, which in turn can affect the therapeutic effect. While these variations may occur due to process control limitations, these variations may also change over time due to the dynamic nature of the process. For example, materials may deposit and accumulate on the walls of the mixing chambers (414, 420, 550, 552) over time, thereby changing the properties of the mixing chambers (414, 420, 550, 552) and thus changing the output from the mixing chambers (414, 420, 550, 552). While some such variations are tolerable to some extent, it may be advantageous to provide quality control features to monitor whether the output of the mixing chambers (414, 420, 550, 552) has deviated beyond tolerance. It may also be desirable for such quality control features to provide such monitoring on the processing chips (111, 200, 500) without the need for fluid to be transferred from the processing chips (111, 200, 500) to an external quality control stage.

[0122] In the context of the encapsulated mRNA particle examples provided above, the output parameters of the variable mixing chambers (414, 420, 550, 552) include the particle size and / or particle size distribution of the encapsulated mRNA particles. In the case where the particle size and / or particle size distribution of the encapsulated mRNA particles from the mixing chambers (414, 420, 550, 552) deviates beyond tolerance, such deviation may indicate excessive accumulation of material on the walls of the mixing chambers (414, 420, 550, 552) and / or other undesirable conditions. Additionally, an unacceptable particle size and / or particle size distribution of the encapsulated mRNA particles may render such encapsulated mRNA particles unavailable for therapy, and thus it may be desirable to immediately stop the output from any mixing chamber (414, 420, 550, 552) that is outputting encapsulated mRNA particles with a particle size and / or particle size distribution that deviates beyond tolerance.

[0123] One method that can be used to measure the particle size and / or particle size distribution of particles (e.g., encapsulated mRNA particles, etc.) is dynamic light scattering (DLS). DLS involves projecting light into a liquid containing the particles, where the particles in the liquid scatter the light. An optical sensor is used to sense the pattern of the light scattered by the particles. As the particles disperse in the liquid due to Brownian motion, this scattering pattern changes over time. Due to Brownian motion, the particles may initially be highly correlated; then ultimately transition to an uncorrelated state. Autocorrelation can be used to track the change in the scattering pattern over time. Since the autocorrelation function is plotted over time, the resulting curve can be analyzed to determine particle diffusion, which can be used to determine the particle size and / or particle size distribution. In some scenarios, the shape of the autocorrelation function curve can indicate whether the encapsulation process performed through one or more mixing chambers (414, 420, 550, 552) was successful (i.e., within tolerance).

[0124] The following examples illustrate how DLS can be incorporated into processing chips (111, 200, 500). Although these examples are provided in the context of encapsulating mRNA particles, the same teachings can be readily applied to other contexts where some other kind of particles are generated by the processing chips (111, 200, 500). Similarly, although the examples are provided in the context of the output from a mixing chamber, the same teachings can be readily applied to the output from other kinds of stages in the processing chips (111, 200, 500). Thus, the following teachings are not necessarily limited to encapsulating mRNA particles or the output from a mixing chamber.

[0125] A. Examples of Processing Chip Configurations for Dynamic Light Scattering

[0126] Figures 9 to 11 An example of a processing chip (800) and a body (900) is shown. The processing chip (800) of this example can be used to generate encapsulated mRNA particles; while the body (900) can be coupled to other optical components as described below to provide a dynamic light scattering stage. The processing chip (800) of this example has a generally square shape, where the body (900) is positioned near a corner of the square shape. This positioning can minimize the encroachment of the body (900) on the upper surface (806) of the processing chip (800), thus leaving space for other components to be positioned on the upper surface (806) of the processing chip (800); and prevent the body (900) from substantially obscuring areas of the processing chip (800) that may need to be visually observed by sensors (105, 160) or a camera (702) etc. Unless otherwise described below, the processing chip (800) can be constructed and operated like the above-described processing chips (111, 200, 500).

[0127] As Figure 10 best seen in, the processing chip (800) of this example includes a plurality of fluid channels (802). Each fluid channel (802) has a corresponding fluid input port (804) such that fluid can be delivered to the fluid channel (802) via the corresponding fluid input port (804). Some of the fluid ports (804) can receive fluid from corresponding vials in a reagent rack (107). Additionally or alternatively, some of the fluid input ports (804) can receive fluid from the corresponding fluid output of another processing chip (111, 200, 500). Alternatively, the fluid input port (804) can receive fluid from any other suitable source.

[0128] The fluid channel (802) leads to a number of mixing components (820) integrated into the processing chip (800). In some forms, all of the mixing components (820) on the processing chip (800) have the same kind of fluid input and are designed to all produce the same kind of fluid output. As Figure 12 Best seen in, each mixing component (820) includes a set of vacuum caps (822), a set of inlet valves (824), and a set of mixing chambers (830, 840). Referring to one mixing component (820) as representative of the other mixing components (820), the mixing component (820) includes a first vacuum cap (822a) that receives fluid from a first fluid channel (802a); a second vacuum cap (822b) that receives fluid from a second fluid channel (802b); and a third vacuum cap (822c) that receives fluid from a third fluid channel (802c). Each fluid channel (802a, 802b, 802c) receives fluid via a corresponding fluid input port (804a, 804b, 804c). A first valve (824a) meters the flow of fluid from the first vacuum cap (822a) to a first channel (826a) leading to a first mixing chamber (830). A second valve (824b) meters the flow of fluid from the second vacuum cap (822b) to an inlet channel (826b) leading to the first mixing chamber (830). The channels (826a, 826b) converge to form an inlet channel (832) leading to the first mixing chamber (830). The fluids from the channels (826a, 826b) are thus mixed together within the first mixing chamber (830).

[0129] A third valve (824c) meters the flow of fluid from the third vacuum cap (822c) to a third channel (826c) leading to a second mixing chamber (840). An outlet channel (834) from the first mixing chamber (830) converges with the third channel (826c) to form an inlet channel (842) leading to the second mixing chamber (840). The fluids from the channels (834, 826c) are thus mixed together within the second mixing chamber (840). The fluid mixed in the second mixing chamber (840) is output through an outlet channel (844).

[0130] In some forms of using hybrid components to provide encapsulation of mRNA, a combination of mRNA and water can be delivered through a first fluid channel (802a), and one or more delivery carrier molecules in ethanol can be delivered through a second fluid channel (802b). In this type of form, the mRNA and the delivery carrier molecules can thus be combined in a first mixing chamber (830) for encapsulation. A diluent (e.g., a citrate-based buffer solution, etc.) can be delivered through a third fluid channel (802c). In this type of form, a second mixing chamber (840) can thus be used to provide pH adjustment. In some variations, the mRNA and water are combined in another mixing chamber (not shown) upstream of the first fluid channel (802a). Similarly, the delivery carrier molecules and ethanol can be combined in another mixing chamber (not shown) upstream of the second fluid channel (802b).

[0131] An additional channel (852) is fluidly coupled to the outlet channel (844) via an opening (850). The channel (852) leads to a fluid output port (853) that can be fluidly coupled to a collection vial (e.g., for storage, etc.) in a reagent rack (107), to another processing chip (111, 200, 500, 800) (e.g., for further processing, etc.), or to any other object. Another valve (854) is positioned downstream of the outlet channel (844) and the opening (850); and upstream of the manifold inlet channel (856). When the valve (854) is in the closed state, the fluid from the outlet channel (844) can only flow through the opening (850) and the channel (852), such that the fluid from the outlet channel (844) will ultimately be delivered out through the fluid output port (853). When the valve (854) is in the open state, at least some of the fluid from the outlet channel (844) can flow through the manifold inlet channel (856). In some variations, another valve (not shown) is placed in the channel (852). In this type of form, this another valve can be switched to the closed state when the valve (854) is in the open state to ensure that the fluid from the outlet channel (844) only flows through the manifold inlet channel (856) and not through the channel (852). Thus, when the valve in the channel (852) is in the closed state, the fluid from the outlet channel (844) can leave without passing through the fluid output port (853). When the valve (854) is in the closed state, this another valve in the channel (852) can be switched to the open state.

[0132] All of the manifold inlet channels (856) of the mixing stages (820) are fluidly coupled to a common manifold channel (858) that ultimately leads to the manifold outlet channel (860). The channels (856, 858, 860) thus cooperate to form a manifold. In this example, an actuatable valve (854) is provided such that at any given time fluid from only one of the manifold inlet channels (856) is conveyed to the channels (858, 860). In other words, during some operating modes of the processing chip (800), only one valve (854) is open at any given moment while the other valves (854) are closed. This ensures that at any given time only the output of one mixing assembly (820) is subjected to DLS testing (described in more detail below). In some scenarios, all of the valves (854) may be closed during certain time periods.

[0133] Referring again to Figure 10 , the processing chip (800) of this example includes three pressure sensing regions (810). A first pressure sensing region (810a) is fluidly coupled to a first fluid channel (802a) such that the first pressure sensing region (810a) can detect the pressure of the fluid within the first fluid channel (802a). A second pressure sensing region (810b) is fluidly coupled to a second fluid channel (802b) such that the second pressure sensing region (810b) can detect the pressure of the fluid within the second fluid channel (802b). A third pressure sensing region (810c) is fluidly coupled to a third fluid channel (802c) such that the third pressure sensing region (810c) can detect the pressure of the fluid within the third fluid channel (802c). The pressure sensing regions (810) can be part of a pressure sensing stage constructed and operated like the pressure sensing stage (700) described above. Although Figure 10 illustrates a set of pressure sensing regions (810) coupled to only one mixing assembly (820), other configurations can be constructed to have the same arrangement of pressure sensing regions (810) for each mixing assembly (820).

[0134] As will be described in more detail below, the pressure sensing stage at the pressure sensing regions (810) can provide pressure data associated with the fluid inlets to the mixing assemblies (820), which can be used to determine the fluid flow rate through the mixing assemblies (820). Such flow rates can be correlated with the particle size and / or particle size distribution data from the DLS stage to determine whether the mixing assemblies (820) are operating within acceptable parameters. In other words, when the DLS data indicates that the particle size and / or particle size distribution of the encapsulated mRNA particles deviates beyond tolerance, the data from the pressure sensing stage at the pressure sensing regions (810) can provide further indication of where within the processing chip (800) a corresponding failure is occurring.

[0135] Figures 13 to 15Shows components of the DLS stage integrated into a processing chip (800). As shown, the manifold outlet channel (860) from the hybrid component (820) leads to the DLS chamber (870). In this example, the fluid from the manifold outlet channel (860) passes through the DLS pre-stage (890) before reaching the inlet channel (872) of the DLS chamber (870). In some configurations, the DLS pre-stage (890) includes a dilution stage that dilutes the fluid before it reaches the DLS chamber (870). Such a dilution stage can be constructed and operated in any suitable manner. Additionally or alternatively, the DLS pre-stage (890) can take any other suitable form and can provide any other suitable function (e.g., ventilation, etc.). In some configurations, the DLS pre-stage (890) is omitted.

[0136] As Figures 14 to 15 Best seen in, the DLS chamber (870) of this example includes a circular upper wall (876), a circular lower wall (878), and a cylindrical side wall (880) extending between the walls (876, 878). The walls (876, 878) and the side wall (880) cooperate to define a hollow interior. The inlet channel (872) provides an inlet for fluid to enter the hollow interior of the DLS chamber through the side wall (880) near the circular lower wall (878). The outlet channel (874) provides an outlet for fluid to leave the hollow interior of the DLS chamber through the side wall (880) near the circular upper wall (876). When viewed from one end of the DLS chamber (870), the channels (872, 874) are oriented generally tangentially with respect to the side wall (880); and are angularly spaced apart from each other by approximately 180 degrees. The cylindrical configuration of the DLS chamber (870) and the vertical positioning and tangential orientation of the channels (872, 874) can provide a sweeping flow of fluid through the interior of the DLS chamber (870), which can help remove bubbles that might otherwise adversely affect the DLS process. In some scenarios, this flow substantially stops within the DLS chamber (870) during the DLS process.

[0137] Although in this example the DLS chamber (870) has a cylindrical configuration (e.g., having a circular cross-sectional shape), the DLS chamber (870) may alternatively have any other suitable configuration. For example, the DLS chamber (870) may have an elliptical cross-sectional shape (when viewed from top to bottom), a pointed-elliptical cross-sectional shape (when viewed from top to bottom), or any other suitable cross-sectional shape rather than the circular cross-sectional shape as shown. Similarly, although the channels (872, 874) are shown as having a generally rectangular cross-sectional shape, the channels (872, 874) may have any other suitable cross-sectional shape. By way of example only, some forms of the channels (872, 874) may taper along the vertical dimension and / or along the horizontal dimension. In some such forms, the wider end of the taper may be located at the DLS chamber (870), while the narrower end of the taper is further away from the DLS chamber (870). Such tapers may be linear and / or curved.

[0138] As described in more detail below, the DLS process includes projecting light into the DLS chamber (870) such that the light will be scattered by particles in the fluid within the DLS chamber (870). The scattered light is detected and processed to establish an autocorrelation curve. In this example, the projected light and the scattered light pass through a region of the processing chip (800) located above the DLS chamber (870). Accordingly, at least this region of the processing chip (800) includes a light-transmissive material (e.g., glass, plastic, silicone, and / or any other suitable material). Such materials may allow the projected light and the scattered light to pass through the region between the upper surface (806) of the processing chip (800) and the DLS chamber (870), and such materials do not cause additional scattering of the light (which could otherwise adversely affect the accuracy of the DLS readings).

[0139] In addition, the region of the processing chip (800) located at or below the circular lower wall (878) and / or other regions of the processing chip (800) may include a light-absorbing material. For example, such materials may be applied to a region of the lower surface (808) of the processing chip (800); or somewhere between the lower surface (808) and the circular lower wall (878) of the DLS chamber (870). Such light-absorbing materials may prevent or minimize ambient light intrusion into the DLS chamber (870) that might otherwise occur. In some forms, the circular lower wall (878) and the sidewall (880) are coated with an opaque material that does not scatter light. As another variant, the material at or below the circular lower wall (878) and / or other regions of the processing chip (800) may be oriented or otherwise configured to reflect light directionally away from the sensing optical fiber (1020) in the body (900)( Figure 16 )). For example, a piece of wedge-shaped smoked glass or the like may be positioned at or below the circular lower wall (878) and / or other regions of the processing chip (800).

[0140] The processing chip (800) of this example further includes a DLS post-stage (892) fluidly coupled to the outlet channel (874) of the DLS chamber (870). In some forms, the DLS post-stage (892) is used in combination with the DLS pre-stage (890) to remove air bubbles from the fluid to be tested in the DLS chamber (870). For example, the fluid can be pumped back and forth between the stages (890, 892) before it can stay in the DLS chamber (870) for the DLS process. Additionally or alternatively, the DLS post-stage (892) can take any other suitable form and can provide any other suitable function (such as ventilation, etc.). In some forms, the DLS post-stage (892) is omitted. The outlet channel (896) is fluidly coupled to the DLS post-stage (892) and leads to the output port (898). The fluid that has passed through the DLS chamber (870) can thus ultimately be conveyed out of the processing chip (800) via the outlet channel (896) and the output port (898). In forms where the DLS post-stage (892) is omitted, the outlet channel (874) of the DLS chamber (870) can be directly fluidly coupled to the output port (898).

[0141] In some forms, the fluid from the output port (898) is conveyed to a waste storage compartment. Such a waste storage compartment can be located in another processing chip (111, 200, 500), in the reagent storage rack (107), or elsewhere. In some other forms, the fluid from the output port (898) is further processed to form a therapeutic composition. Such further processing can include conveying the fluid to another processing chip (111, 200, 500, 800), to a vial in the reagent storage rack (107), or elsewhere. As another option, the fluid from the output port (898) can be used for some other quality control tests (such as on another processing chip (111, 200, 500, 800) or elsewhere). Alternatively, the fluid from the output port (898) can be processed in any other suitable manner.

[0142] B. Examples of Optical Components for Dynamic Light Scattering

[0143] Figures 16 to 20 The body (900) is shown in more detail. The body (900) of this example includes an elongated vertical portion (910), a lateral portion (920), and a mounting flange (930). The vertical portion (910) defines an upper channel (912), a lens placement area (915) positioned below the upper channel (912), and a closed focusing volume (916) positioned below the lens placement area (915). As Figures 16 to 17As shown, the upper channel (912) is sized to receive a collimator (1000); and the lens placement area (915) is sized to receive a focusing lens (1002). The clamping flange (914) extends laterally from the vertical portion (910). When the collimator (1000) is inserted into the upper channel (912), a screw or other component can be secured to the clamping flange (914) such that the vertical portion (910) can thereby clamp the collimator (1000) and hold the collimator (1000) firmly in the upper channel (912). Alternatively, the collimator (1000) can be held in any other suitable manner. The focusing lens (1002) is captured between the collimator (1000) and the closed focusing volume (916). The closed focusing volume (916) defines a tapered optical path that tapers toward an opening (918) formed through the lower surface (902) of the body (900). In this example, the sidewall (917) of the closed focusing volume (916) includes light-absorbing material. As described in more detail below, the vertical portion (910) projects light along a first axis (A 1 ) through the opening (918).

[0144] The lateral portion (920) extends laterally from the vertical portion (910). The lateral portion (920) defines an upper channel (922) and a lower channel (928), where a filter slot (926) interrupts the lower channel (928). As Figure 16 shown, the upper channel (922) is sized to receive a sensing optical fiber (1020). In some configurations, the sensing optical fiber (1020) is a multimode optical fiber. In some such configurations, the sensing optical fiber (1020) has a core diameter in the range of from about 50 μm to about 400 μm; or about 50 μm. In some other configurations, the sensing optical fiber (1020) is a single-mode optical fiber. In some such configurations, the sensing optical fiber (1020) has a core diameter in the range of from about 2 μm to about 11 μm; or about 5 μm.

[0145] When the sensing optical fiber (1020) is inserted into the upper channel (922), a screw (924) can be inserted into the transverse portion (920) and tightened so that the transverse portion (920) can thereby fix the sensing optical fiber (1020) and firmly hold the sensing optical fiber (1020) in the upper channel (922). In this example, a ferrule (not shown) is positioned around the region of the sensing optical fiber (1020) inserted into the transverse portion (920) such that the transverse portion (920) effectively clamps the sensing optical fiber (1020) via the ferrule. Alternatively, the sensing optical fiber (1020) can be held in any other suitable manner. The lower channel (928) defines a cylindrical optical path leading to an opening (929) formed through the lower surface (902) of the body (900). In this example, the lower channel (928) includes a light-absorbing material. As described in more detail below, the transverse portion (920) conveys light received through the opening (929) along a second axis (A 2 )). As best visible in Figures 19 to 20 , the openings (918, 929) are spaced apart from each other in this example such that the openings (918, 929) do not overlap each other.

[0146] As Figure 17 shown, a filter (1004) can be inserted into the filter slot (926). The filter (1004) can thus filter the light received through the opening (929) and the lower channel (928) before further conveying such light to the sensing optical fiber (1020) in the upper channel (922). As part of this filtering, the filter (1004) can help remove the wavelengths of light associated with ambient illumination. The filter (1004) can thus be tuned to match (or otherwise accommodate) the wavelength of the light emitted through the source optical fiber (1010) (i.e., the incident light), as described in more detail below. In some variations, the filter (1004) can also be used to detect fluorescence (e.g., by filtering the wavelength of the source light). With reference to the teachings herein, various suitable forms and configurations for the filter (1004) will be apparent to those skilled in the art.

[0147] The mounting flange (930) extends laterally from the vertical portion (910) adjacent to the transverse portion (920). The mounting flange (930) is configured to receive a screw (930)( Figure 16 ) and thereby fix the body (900) to a fixing device (e.g., the seating mount (154) of the base (180)) such that the body (900) can be firmly fixed relative to the base (180). In other words, in this example, the body (900) is not fixed to the processing chip (800). Instead, as Figure 11As shown, the main body (900) can be fixed via the mounting flange (930) such that the lower surface (902) of the main body (900) is parallel to the upper surface (806) of the processing chip (800); and such that the lower surface (902) is spaced apart from the upper surface (806) by a gap distance (d). By way of example only, the range of the gap distance (d) can be from about 0 mm to about 0.5 mm; or can be about 0 mm. Alternatively, any other suitable gap distance (d) can be used. In the form where the gap distance (d) is 0 mm, such that the lower surface (902) of the main body (900) contacts the upper surface (806) of the processing chip (800), one or both of the lower surface (902) or the upper surface (806) can include complementary surface features (e.g., protrusions and / or recesses) that assist in properly registering the position of the main body (900) relative to the processing chip (800).

[0148] When the main body (900) is fixed at a gap distance (d) from the upper surface (806) of the processing chip (800), the axis (A 1 、A 2 ) converges at a convergence point (CP) located within the DLS chamber (870). The position of the convergence point (CP) can be indicated by the angle (θ) formed by the axes (A 1 、A 2 ). Also by way of example only, the range of the angle (θ) formed by the axes (A 1 、A 2 ) can be from about 10 degrees to about 90 degrees; or the range is from about 10 degrees to about 45 degrees; or is about 28.8 degrees. Also by way of example only, the convergence point (CP) can be positioned at a distance ranging from about 1 mm to about 5 mm; or about 1.2 mm from the lower surface (902).

[0149] As Figure 16As shown, one end of the source optical fiber (1010) can be coupled to the collimator (1000). Another end of the source optical fiber (1010) can be coupled to the coherent light source (1012). By way of example only, the light source (1012) can include a single-frequency laser capable of operating to emit light at a certain wavelength at which the processing chip (800) is optically transmissive. The light source (1012) can be mounted on the base (180) or otherwise integrated into the system (100). In some forms, the light source (1012) communicates with the controller (121). For example, the controller (121) is capable of operating to selectively activate the light source (121) and tune the intensity of the light generated by the light source (121). Additionally or alternatively, the controller (121) can receive feedback related to the operation of the light source (121). In some other forms, the light source (121) is controlled and / or monitored independently of the controller (121). In this example, the light generated by the light source (1012) is transmitted along the source optical fiber (1010) to the collimator (1000). In terms of the safety issues regarding the collimated light to a human operator, it may be advantageous to position the collimator (1000) in the body (900) where the collimated light will not reach the eyes of the human operator. The collimated light from the collimator (1000) further passes through the focusing lens (1002), through the closed focusing volume (916), and then exits through the opening (918). The projected light ultimately reaches the DLS chamber (970) as described above.

[0150] Also as Figure 16 shown, one end of the sensing optical fiber (1020) can be fixed within the upper channel (922), while another end of the sensing optical fiber (1020) can be coupled to the photon counter (1022), and the photon counter can be further coupled to the autocorrelator (1024). The photon counter (1022) and the autocorrelator (1024) can take any suitable form, which will be apparent to those skilled in the art in view of the teachings herein. When the light projected through the opening (918) is scattered by the particles in the fluid in the DLS chamber (970), the scattered light is transmitted through the opening (929), the lower channel (928), and the filter (1004), and ultimately reaches the sensing optical fiber (1020). The sensing optical fiber (1020) also transmits the light to the photon counter (1022), which counts the photons and transmits the corresponding photon count data to the autocorrelator (1024). The autocorrelator (1024) performs an autocorrelation function on the data to generate such as Figures 21 to 22Those autocorrelation curves as shown and described in more detail below. The autocorrelator (1024) can be further coupled to the controller (121) such that the controller (121) can process the autocorrelation data from the autocorrelator (1024) and use at least such data to perform algorithms. Examples of processes that can be performed using at least the autocorrelation data will be described in more detail below. Although Figure 16 the autocorrelator (1024) and the controller (121) are depicted as separate components, some variations of the controller (121) can directly integrate the autocorrelation module or otherwise provide the autocorrelation function. In other words, some forms may not have the autocorrelator (1024) and the controller (121) as separate components.

[0151] Figure 21 A diagram (1100) is shown that depicts an example of plotting autocorrelation curves (1110, 1120) that can be formed using the DLS stage as described above. In Figure 21 the example shown, the autocorrelation curve (1110) represents data obtained by using a multimode fiber for the sensing fiber (1020) to detect DLS in a fluid containing light-scattering beads. The autocorrelation curve (1120) represents data obtained by using a multimode fiber for the sensing fiber (1020) to detect DLS in water (i.e., as a control). In this example, the autocorrelation curve (1110) has a y-intercept of approximately 0.009. To the extent that the autocorrelation curve (1110) is somewhat unstable or noisy during a very early stage of this time period, the autocorrelation curve (1110) quickly turns into a relatively smooth curve with little noise.

[0152] Figure 22 A diagram (1200) is shown that depicts another example of plotting autocorrelation curves (1210, 1220) that can be formed using the DLS stage as described above. In Figure 22 the example shown, the autocorrelation curve (1210) represents data obtained by using a single-mode fiber for the sensing fiber (1020) to detect DLS in a fluid containing light-scattering beads. The autocorrelation curve (1220) represents data obtained by using a multimode fiber for the sensing fiber (1020) to detect DLS in a fluid containing light-scattering beads. In other words, Figure 22 the autocorrelation curve (1220) in Figure 21 is similar to the autocorrelation curve (1110) in

[0153] According to Figures 21 to 22 the illustrated diagrams (1100, 1200) shown, acceptable results can be produced using a multimode fiber for the sensing fiber (1020) or using a single-mode fiber for the sensing fiber (1020). The multimode fiber can still obtain a useful and reliable autocorrelation curve (1110) in that the single-mode fiber can generate a much higher y-intercept in its autocorrelation curve (1210).

[0154] C. Examples of Methods Using the Dynamic Light Scattering Stage

[0155] In a method of operating a processing chip (800), various fluid components can be conveyed through a fluid input port (804) and a fluid channel (802) to reach a mixing assembly (820). Such fluid components can include mRNA, surfactants, pH-adjusting buffers, ethanol, water, and / or other components. The output of each mixing assembly (820) can include encapsulated mRNA particles. A valve (854) can be operated to sequentially direct the output of each mixing assembly (820) via channels (858, 860) towards a DLS stage. When the sample volume of the fluid output of one mixing assembly (820) reaches a DLS chamber (870), light from a light source (1012) can be emitted into the DLS chamber (870) via a source fiber (1010) and associated components of a body (900). In some forms, the flow of fluid in the DLS chamber (870) stops during the DLS process such that once the fluid in the DLS chamber (870) has reached a substantially stationary state, light from the light source (1012) is emitted into the DLS chamber (870). In some other forms, light from the light source (1012) is emitted into the DLS chamber (870) while the fluid in the DLS chamber (870) is in a flowing state.

[0156] When light from the light source (1012) is emitted into the DLS chamber (870), regardless of whether the fluid in the DLS chamber (870) is stationary or flowing, the light can be scattered by particles in the sample volume in the DLS chamber (870). The scattered light can be conveyed through a sensing fiber (1020) and associated components of the body (900) and ultimately reach a photon counter (1022). An autocorrelator (1024) can process the corresponding data from the photon counter (1022) and generate a similar to Figures 21 to 22The autocorrelation curve shown. The autocorrelation curve can be compared with a baseline curve to determine whether the autocorrelation curve is within tolerance. In some embodiments, this comparison is performed by a controller (121). Determining whether the autocorrelation curve is within tolerance can include determining whether the y-intercept of the curve is at an appropriate level, whether the curve follows a path within a tolerable deviation from the path of the baseline curve, the degree to which the curve fits a certain model (e.g., cumulants, etc.), whether the curve drops to a value close to zero, the autocorrelation time at which a transition occurs, and / or whether the autocorrelation curve meets other criteria.

[0157] If the autocorrelation curve is within tolerance, this can indicate that the particle size and / or particle size distribution of particles (e.g., encapsulated mRNA particles, etc.) in the DLS chamber (870) is appropriate; and the mixing assembly (820) that produces the sample volume in the DLS chamber (870) is operating properly. The processing chip (800) can then continue to deliver the output of the mixing assembly (820) to a subsequent stage (e.g., a storage vial in the reagent storage rack (107), another processing chip (111, 200, 500), etc.). If the autocorrelation curve is outside of tolerance, this can indicate that the particle size and / or particle size distribution of particles (e.g., encapsulated mRNA particles, etc.) in the DLS chamber (870) is inappropriate; and the mixing assembly (820) that produces the sample volume in the DLS chamber (870) is operating improperly.

[0158] If the mixing assembly (820) fails at the DLS stage (i.e., DLS data indicates that the particle size and / or particle size distribution of the output from the mixing assembly (820) is outside of tolerance), then the mixing assembly (820) can be effectively shut down. The mixing assembly (820) can be effectively shut down by no additional fluid being conveyed through the fluid input port (804) upstream of the mixing assembly (820). Additionally, no fluid will be conveyed from the fluid output port 853 downstream of the mixing assembly 820 that has been effectively shut down. In some embodiments, the controller (121) executes an algorithm to ensure that no additional fluid is conveyed through the fluid input port (804) upstream of the mixing assembly (820) that fails at the DLS stage by automatically preventing fluid communication from any fluid source (e.g., a vial in the reagent storage rack (107), the output of another processing chip (111, 200, 500), etc.), which would otherwise supply fluid to such a fluid input port (804).

[0159] Each DLS measurement process can be performed for any suitable duration. For example, in some embodiments, the process of emitting light into the fluid and collecting the scattered light can be performed for a duration ranging from about 1 second to about 60 seconds starting from the time the fluid in the sample volume reaches the DLS chamber (870); or the duration can be about 10 seconds. After performing DLS on the fluid in the sample volume, the fluid in the sample volume can be processed in any suitable manner. For example, if DLS indicates that the particle size and / or particle size distribution in the fluid is appropriate, the sample volume can be further transferred to a vial in the reagent rack (107), to another processing chip (111, 200, 500) for further processing, to a waste reservoir, or elsewhere. If DLS indicates that the particle size and / or particle size distribution in the fluid is inappropriate, the sample volume can be further transferred to a waste reservoir or elsewhere. In some embodiments, the DLS measurement can be performed several times (e.g., a range of 1 to 20 iterations); or more specifically 10 times). Such repeated measurements can be used to collect statistical data, capture time-varying conditions, etc.

[0160] Regardless of whether the output of the mixing assembly (820) has produced an acceptable or unacceptable particle size and / or particle size distribution, after the output of the first mixing assembly (820) has been analyzed in the DLS stage, the processing chip (800) can operate the valve (854) to transfer the output of the next mixing assembly (820) to the DLS stage. This sequence can continue until the output of each mixing assembly (820) has been analyzed in the DLS stage. Once the output of the last mixing assembly (820) has been analyzed in the DLS stage, the process can start from the first mixing assembly (820) and continue to execute this sequence throughout the duration of the operation of the processing chip (800). In the case where any mixing assembly (820) stops working due to its output failure in the DLS stage, such mixing assembly (820) can be ignored as the processing chip (800) repeats the sequence of testing the output of the mixing assembly (820) in the DLS stage.

[0161] In some cases, it may be desirable to provide some kind of spacer or purge fluid through the DLS stage between the outputs of the mixing assemblies (820). For example, air bubbles can be transferred through the DLS chamber (870) between the outputs of the mixing assemblies (820). Additionally or alternatively, a predetermined water or some other liquid can be transferred through the DLS chamber (870) between the outputs of the mixing assemblies (820). In either case, the air, water, or other fluid can help clear any particles that may otherwise remain in the DLS chamber (870) before the output of the next mixing assembly (820) reaches the DLS chamber (870).

[0162] In a type where the processing chip (800) includes a pressure sensing region (810) that is part of a pressure sensing stage constructed and operating like the pressure sensing stage (700) described above, such a pressure sensing stage can be used in combination with the DLS stage in a variety of ways. In some types, the controller (121) initially monitors only the pressure data from the pressure sensing stage and does not activate the DLS stage until the pressure data indicates that the pressure level in one or more fluid channels (802) is outside of tolerance. For example, if the pressure level in the fluid channel (802) exceeds a predetermined threshold, the elevated pressure level may indicate that a blockage is forming (or has formed) in the mixing assembly (820) fed by the fluid channel (802). If this occurs, the controller (121) can open the valve (854) downstream of the mixing assembly (820) to direct the output from the mixing assembly (820) to the DLS stage for testing. Once the output from the mixing assembly (820) reaches the DLS chamber (870), the DLS stage can be used to determine whether the particle size and / or particle size distribution of the particles from the mixing assembly (820) is appropriate. If so, the DLS stage can be used to continue monitoring the output of the mixing assembly (820) to determine whether and when the particle size and / or particle size distribution falls outside of tolerance (at which point the mixing assembly (820) can effectively be shut down).

[0163] In some other forms in which the processing chip (800) includes a pressure sensing region (810) that is part of a pressure sensing stage constructed and operating like the pressure sensing stage (700) described above, the controller (121) can observe pressure data in response to the DLS stage detecting a particle size and / or particle size distribution that falls outside of the tolerance. In other words, if the DLS stage determines that the output from the mixing assembly (820) includes an unacceptable particle size and / or particle size distribution, the controller (121) can observe the fluid pressure in the fluid channels (802) that serve as the inlet to the mixing assembly (820) and determine whether any of these fluid channels (802) has a fluid pressure that falls outside of the tolerance. For example, if the DLS stage determines that the output from the mixing assembly (820) includes an unacceptable particle size and / or particle size distribution, the controller (121) can determine that one of the fluid channels (802) feeding the mixing assembly (820) has an unacceptably high fluid pressure; and this can indicate that a blockage is forming (or has formed) in the mixing assembly (820). The mixing assembly (820) can then be effectively shut down and / or treated (e.g., chemically treated, etc.) to remove the blockage, such that only other mixing assemblies (820) on the processing chip (800) are used thereafter (or until the treatment to remove the blockage is complete). In the case where the DLS stage determines that the output from the mixing assembly (820) includes an unacceptable particle size and / or particle size distribution and the fluid pressure in the fluid channels (802) feeding the mixing assembly (820) is not abnormal, then this can indicate that there is some other kind of problem (e.g., chemical composition change, etc.) and appropriate remedial measures can be taken.

[0164] While the foregoing examples provide a sequence of DLS measurements followed by fluid pressure measurements (where the fluid pressure measurements are triggered by a particular particle size and / or particle size distribution measurement), or a sequence of fluid pressure measurements followed by DLS measurements (where the DLS measurements are triggered by a particular fluid pressure measurement), some other variations can provide pressure measurements and DLS measurements in parallel. For example, the controller (121) can continuously monitor pressure measurements and DLS measurements; and when a combination of pressure measurements and particle size and / or particle size distribution measurements meets a particular criterion, determine that the mixing assembly (820) should be shut down.

[0165] Regardless of whether and how the fluid pressure measurement and the DLS measurement are performed in a specific sequence (i.e., first the pressure measurement is performed, then the DLS measurement; first the DLS measurement is performed, then the fluid pressure measurement; or the pressure measurement and the DLS measurement are performed in parallel), the controller (121) can provide a certain degree of artificial intelligence when learning from the correlation between the DLS measurement results and the fluid pressure measurement results. For example, the controller (121) can learn which fluid pressure levels tend to correspond to points where the DLS measurement results indicate unacceptable particle size and / or particle size distribution; and the controller (121) can thus, before the mixing component (820) produces an unacceptable particle size and / or particle size distribution, based on the fluid pressure value associated with the mixing component (820), shut down the mixing component (820) in advance. In other words, the controller (121) can use the correlation between at least the sensed fluid pressure and the detected unacceptable particle size and / or particle size distribution to adjust the fluid pressure threshold.

[0166] Regardless of whether the pressure data from the pressure sensing area (810) is related to the DLS data or how it is related, in some embodiments, the pressure data from the pressure sensing area (810) can also be used to change the input via the fluid port (804). For example, if the pressure data at the pressure sensing area (810a) indicates that the pressure of the fluid in the fluid channel (802a) is higher than a certain value, the controller (121) can activate the pressure source (117) to reduce the pressure of the fluid being transmitted to the port (804a). Similarly, if the pressure data at the pressure sensing area (810a) indicates that the pressure of the fluid in the fluid channel (802a) is lower than a certain value, the controller (121) can activate the pressure source (117) to increase the pressure of the fluid being transmitted to the port (804a). The pressure data at the pressure sensing area (810b) can be used similarly to track the pressure of the fluid in the fluid channel (802b); and such pressure data can be used to change the pressure of the fluid being transmitted to the fluid input port (804b). Likewise, the pressure data at the pressure sensing area (810c) can be used similarly to track the pressure of the fluid in the fluid channel (802c); and such pressure data can be used to change the pressure of the fluid being transmitted to the fluid input port (804c). The pressure sensing level at the pressure sensing area (810) can thus be used to provide a feedback loop, enabling real-time adjustment of the pressure of the fluid being transmitted to the fluid input port (804) to achieve consistency of the desired pressure value.

[0167] In some scenarios, pressure data from a pressure sensing region (810) is compared with the pressure of the fluid in the corresponding fluid channel upstream of the port (804) provided by a pressure source (117) to determine whether the pressure data from the pressure sensing region (810) deviates acceptably from the pressure in the corresponding fluid channel upstream of the port (804). In such cases, some deviation can be expected, such as the fluid pressure in the pressure sensing region (810) being lower than the fluid pressure in the corresponding fluid channel upstream of the port (804). Similarly, the pressure data from the pressure sensing region (810) can be compared with the fluid pressure in the corresponding fluid channel upstream of the port (804) provided by the pressure source (117) to determine the flow rate of the fluid flowing through the fluid channel (802). Such flow rate data is particularly useful in scenarios where DLS is performed on flowing rather than stationary fluids. Examples of performing DLS on flowing fluids are described in more detail below.

[0168] As another example of how the pressure sensing level at the pressure sensing region (810) can be used to provide a feedback loop that enables real-time adjustment of the pressure of the fluid being delivered to the fluid input port (804), the pressure data from the pressure sensing regions (810a, 810b, 810c) associated with a given mixing assembly (820) can be evaluated to ensure that the fluid pressures along the channel (802) are properly balanced with each other. Similarly, in cases where the pressure data from the pressure sensing regions (810a, 810b, 810c) is compared with the pressure data from the corresponding fluid channels upstream of the fluid input port (804) to determine the flow rate of the fluid flowing through the channel (802), the flow rate of the fluid flowing through the channel (802) can be evaluated to ensure that the flow rates flowing through the channel (804) are properly balanced with each other. In cases where the fluid pressures and / or flow rates in the channel (802) are not properly balanced with each other, the controller (121) can activate the pressure source (117) to increase or decrease the pressure of the fluid being delivered to any fluid input port (804) where such adjustment is reasonable.

[0169] The foregoing examples of pressure data used in a real-time feedback loop refer to pressure regulation via the pressure source (117) to increase or decrease the pressure of the fluid being delivered to the fluid input port (804). In some variants, the fluid pressure is generated or otherwise controlled via peristaltic pumping performed on the processing chip (800) (e.g., as described above in the context of the processing chips (111, 200, 500)). In such variants, the controller (121) can adjust the peristaltic pumping action on the processing chip (800) in response to the pressure data from the pressure sensing region (810).

[0170] D. Examples of Processing Chips with Multiple Dynamic Light Scattering Stages

[0171] While Figures 9 to 11 the arrangement is shown with only one DLS stage being used with the processing chip (800), it may be desirable to provide an arrangement in which the processing chip (800) includes a plurality of DLS stages. Figures 23 to 24 An example of such an alternative arrangement is shown. In this example, a body (900) is positioned near a corner of the processing chip (800), similar to Figures 9 to 11 the arrangement shown. However, unlike Figures 9 to 11 the arrangement shown, Figures 23 to 24 the arrangement shown also includes a number of additional bodies (901), each additional body (901) being positioned above a respective mixing component (820). These additional bodies (901) may be constructed and operate like the body (900) described above. In Figures 23 to 24 the arrangement shown, each body (900, 901) may form part of a corresponding DLS stage as described above. Thus, in Figures 23 to 24 the arrangement shown, each mixing component (820) has an associated DLS stage.

[0172] In this example, each body (901) is positioned above a respective outlet channel (844) of the mixing component (820) associated with the body (901). Thus, each DLS stage associated with each body (901) can be used to measure the particle size and / or particle size distribution of the particles in the direct output of each mixing component (820). In some forms of this arrangement, when the fluid is in a flowing state, the DLS stage associated with the body (901) measures the particle size and / or particle size distribution of the particles in the fluid; while when the fluid is in a stationary state, the DLS stage associated with the body (900) measures the particle size and / or particle size distribution of the particles in the fluid. In other words, the DLS stage associated with the body (901) can be regarded as providing a dynamic fluid test, while the DLS stage associated with the body (900) can be regarded as providing a static fluid test.

[0173] In Figures 23 to 24In some forms of the arrangement shown, the DLS stage associated with the body (901) constantly (i.e., as the fluid continues to flow through the corresponding outlet channel (844)) measures the particle size and / or particle size distribution of the particles in the fluid; while the DLS stage associated with the body (900) only temporarily measures the particle size and / or particle size distribution of the particles in the fluid. For example, only when the fluid pressure data and / or DLS data (from the DLS stage associated with the body (901)) indicate that there may be a problem with a particular mixing component (820), a sample volume of the fluid can be transferred to the DLS stage associated with the body (900). In this case, the output of the particular mixing component (820) can be sent to the DLS stage associated with the body (900) to determine whether the particle size and / or particle size distribution of the particles produced by the mixing component (820) is within tolerance.

[0174] As another variant, in addition to the DLS stage associated with the body (901) operating continuously, the DLS stage associated with the body (900) can also operate continuously, and the output from the mixing component (820) is sequentially transferred to this DLS stage. In such a scenario, the DLS data from the stage associated with the body (900) can be correlated with the DLS data from the stage associated with the body (901) corresponding to the mixing component (820), and the output of this mixing component is in the DLS chamber (870). For example, the autocorrelation curves from these DLS stages can be compared with each other; and the controller (121) can determine whether the deviation between these curves is within a predetermined tolerance.

[0175] In addition to providing DLS sensing as described above, the foregoing arrangement can also provide laser Doppler velocimetry. Laser Doppler velocimetry can be used to help determine the flow conditions. As another variant, a tunable electric field can be used to determine the ζ potential indicating the particle charge state. Such particle charge data can indicate the degree of mRNA presence in the particles, since mRNA can have a characteristic charge.

[0176] IV. Examples of Viscosity Sensing via Dynamic Light Scattering

[0177] As described above, it may be desirable to monitor the particle size and / or particle size distribution of the encapsulated mRNA particles during the process of forming the encapsulated mRNA particles via the processing chip (800), because the particle size and / or particle size distribution of the encapsulated mRNA particles may tend to change as the processing chip (800) is used to form the encapsulated mRNA particles. It may be desirable to determine and monitor the viscosity of the solution carrying the encapsulated mRNA particles, because such viscosity may also tend to change as the processing chip (800) is used to form the encapsulated mRNA particles, and the initial viscosity of the fluid may be unknown. For example, some solutions may include a combination of mRNA and water (e.g., introduced via fluid channel (802a)), one or more delivery carrier molecules in ethanol (e.g., introduced via fluid channel (802b)), and a diluent or buffer (e.g., introduced via fluid channel (802c)). A mixture of ethanol and water can produce significant viscosity changes and may thus be very sensitive to changes in ethanol concentration.

[0178] As described in more detail below, a DLS stage as described above can be used to monitor the viscosity of the solution. In addition, the same DLS stage used to monitor the particle size and / or particle size distribution of the encapsulated mRNA particles can also be used to monitor the viscosity of the solution carrying the encapsulated mRNA particles.

[0179] In some scenarios, it may be important to determine and monitor the viscosity of the solution, because changes in solution viscosity may tend to produce measurement effects comparable to those produced by changes in the particle size of the encapsulated mRNA in the solution. This can be observed through the following equation (I):

[0180] (I)

[0181]

[0182] where "d" is the particle diameter of the encapsulated mRNA,

[0183] "λ" is the wavelength of the laser emitted by the light source (1012),

[0184] "v" is the viscosity of the solution,

[0185] "Γ" is the autocorrelation curve fitting value (i.e., the value determined by fitting to the autocorrelation curve),

[0186] "T" is the temperature of the solution,

[0187] "n" is the refractive index of the solution,

[0188] "θ" is the scattering measurement angle formed by the axis (A 1 、A 2 ) of the body (900) in the measurement medium (e.g., liquid or gas), and

[0189] "k" B is the Boltzmann constant.

[0190] Accordingly, to obtain reliable measurements of the particle size and / or particle size distribution of mRNA particles encapsulated within a solution, it may be desirable to obtain a measurement of the solution viscosity. The following description provides examples of how the DLS stage described above can be used to measure and monitor the viscosity of a solution carrying encapsulated mRNA particles and, simultaneously, measure and monitor the particle size and / or particle size distribution of those mRNA particles in the solution. Each example described below provides for the measurement and monitoring of the solution viscosity without any contact between the sensor and the solution being measured.

[0191] Although the following examples describe methods for determining the solution viscosity, the particle size of particles in the solution, and the particle size distribution of particles in the solution, the same methods can be used to determine other parameters, such as refractive index or temperature. Such parameters can be performed using Equation (I) in combination with the data collected as described below.

[0192] A. Examples of Processing Chip Features for Providing Solution Viscosity Measurements

[0193] As referred to above Figure 13 it has been described that the processing chip (800) can include a DLS front stage (890) and a DLS back stage (892), wherein the DLS chamber (870) is positioned in the fluid path between the DLS front stage (890) and the DLS back stage (892). Figure 25 An example of components of a measurement stage (1300) that can be used to effectively form the DLS front stage (890) and the DLS back stage (892) is shown. Accordingly, it should be understood that Figure 25 the measurement stage (1300) can be incorporated into a processing chip similar to the processing chip (800). In this example, the entire set of features of the measurement stage (1300) described below can effectively replace the combination of the DLS front stage (890), the DLS chamber (870), the channels (872, 874), and the DLS back stage (892).

[0194] The measurement stage (1300) is in fluid communication with a sample inlet channel (1302), a test fluid inlet channel (1304), and an outlet channel (1306). In the context of the processing chip (800), the sample inlet channel (1302) can be considered analogous to the manifold outlet channel (860) such that the sample input channel (1302) receives an aliquot (i.e., a sample volume) of a solution (such as output from the mixing assembly (820)) carrying encapsulated mRNA particles. The test fluid inlet channel (1304) is in fluid communication with a test fluid source (not shown). As described in more detail below, the test fluid can include a fluid with beads, a diluent without beads (e.g., buffers and water, etc.), and / or any other suitable type of fluid. The outlet channel (1306) can be considered analogous to the outlet channel (896) such that the solution can exit the measurement stage (1300) via the outlet channel (1306). By way of example only, the outlet channel (1306) can ultimately lead to a waste storage compartment, some other stage that uses the solution to form a therapeutic composition, or some other component.

[0195] The measurement stage (1300) of this example further includes a first valve (1310), a first channel (1312), a first pump (1314), a second valve (1316), a second channel (1318), a third channel (1320), a mixing chamber (1322), a DLS chamber (1324), a fourth channel (1326), a third valve (1328), a fifth channel (1330), a second pump (1332), a sixth channel (1334), and a fourth valve (1336). The valves (1310, 1316, 1328, 1336) of this example can be constructed and operated like the valves (824, 854) described above. Thus, the valves (1310, 1316, 1328, 1336) can drive the elastic layer (e.g., such as the elastic layer (302)) of the processing chip (800) to transition between an open state and a closed state by selectively applying pressure (or releasing pressure on the elastic layer) to the elastic layer in the regions of the processing chip (800) corresponding to the valves (1310, 1316, 1328, 1336). The valves (1310, 1316, 1328, 1336) can thus be used to selectively block or allow fluid flow. The pumps (1314, 1332) can also be operated by selectively applying pressure to the elastic layer (e.g., such as the elastic layer (302)) of the processing chip (800) in the regions of the processing chip (800) corresponding to the pumps (1314, 1332), thereby driving fluid flow through peristaltic pumping action.

[0196] The first valve (1310) is inserted between the sample inlet channel (1302) and the first channel (1312) such that the first valve (1310) can be operated to selectively block or allow fluid flow between the sample inlet channel (1302) and the first channel (1312). The first channel (1312) is also in fluid communication with the first pump (1314). The second valve (1316) is inserted between the test fluid inlet channel (1304) and the second channel (1318) such that the second valve (1316) can be operated to selectively block or allow fluid flow between the test fluid inlet channel (1304) and the second channel (1318). The second channel (1318) is also in fluid communication with the first pump (1314). The first pump (1314) is also in fluid communication with the third channel (1320). The first pump (1314) can be operated to pump fluid from the sample inlet channel (1302) and / or the test fluid inlet channel (1304) towards the third channel (1320), depending on the state of each respective valve (1310, 1316).

[0197] The third channel (1320) is also in fluid communication with the mixing chamber (1322). The mixing chamber (1322) is configured to mix fluids from the sample inlet channel (1302) and the test fluid inlet channel (1304), as pumped into the mixing chamber (1322) via the first pump (1314). By way of example only, the mixing chamber (1322) can be constructed and operated like any mixing chamber described herein. Alternatively, the mixing chamber (1322) can be constructed and operated in any other suitable manner.

[0198] The mixing chamber (1322) leads directly to the DLS chamber (1324). The DLS chamber (1324) can be constructed and operated like the DLS chamber (870) described above. Accordingly, the body (900) can be positioned adjacent to the DLS chamber (1324) such that the source optical fiber (1010) can emit light into the DLS chamber (1324); and such that the sensing optical fiber (1020) can receive light scattered from particles in the fluid in the DLS chamber (1324) as described above. The scattered light can reach the photon counter (1022), and the corresponding data can be processed by the autocorrelator (1024), as described above and as described in more detail below.

[0199] The DLS chamber (1324) is also in fluid communication with a fourth channel (1326) leading to a third valve (1328). The third valve (1328) is also in fluid communication with a fifth channel (1330). The third valve (1328) can thus be operated to selectively block or allow fluid flow between the fourth channel (1326) and the fifth channel (1330). The fifth channel (1330) is also in fluid communication with a second pump (1332). The second pump (1332) is also in fluid communication with a sixth channel (1334). The second pump (1332) can be operated to pump fluid from the fifth channel (1330) towards the sixth channel (1334). The second pump (1332) can also be operated to pump fluid back towards the fifth channel (1330).

[0200] The sixth channel (1334) is also in fluid communication with a fourth valve (1336). The fourth valve (1336) is also in fluid communication with an outlet channel (1306), such that the fourth valve (1336) can be operated to selectively block or allow fluid flow between the sixth channel (1334) and the outlet channel (1306).

[0201] From the foregoing, it should be understood that the pumps (1314, 1332) can drive fluid towards and away from the DLS chamber (1324) from two separate sources. Additionally, the valves (1310, 1316, 1328, 1336) can selectively block fluid flow in different respective regions of the processing chip (800) in conjunction with the operation of the pumps (1314, 1332). How the measurement stage (1300) can be used in combination with Figures 9 to 11 and Figures 16 to 20 the DLS components shown to sense viscosity, particle size, and particle size density will be described in more detail below.

[0202] B. Examples of Viscosity Sensing Using Added Beads

[0203] In some scenarios, the viscosity of a solution can be measured by introducing known beads into the solution and then performing DLS on the solution containing the beads. In this context, "known beads" includes beads having a known diameter. As used herein, the term "bead" should not be understood to be necessarily limited to a spherical structure that is made. For example, "bead" can include other calibrants, such as calibration molecules of known particle size, etc. An example of a calibration bead in the form of a calibration molecule is dextran. Alternatively, any other suitable type of calibration molecule (or other calibrant) can be used.

[0204] Figure 26 An example of a process of using beads via Figure 25 the measurement stage (1300) shown is illustrated. In this example, a solution containing beads and a solution containing encapsulated mRNA particles are delivered to the measurement stage (1300), as Figure 26as shown by the box (1400). This includes opening the first valve (1310) and driving an aliquot of the solution containing encapsulated mRNA particles towards the first pump (1314) via channels (1302, 1312). This also includes opening the second valve (1316) and driving a volume of the solution containing beads towards the first pump (1314) via channels (1304, 1318). In this example, the beads in the solution containing beads have a known particle size; while the encapsulated mRNA particles in the solution containing encapsulated mRNA particles have an unknown particle size and the solution containing encapsulated mRNA particles has an unknown viscosity.

[0205] In some scenarios, the solution containing encapsulated mRNA particles and the solution containing beads are driven towards the first pump (1314) simultaneously. In some other scenarios, the solution containing encapsulated mRNA particles and the solution containing beads are driven towards the first pump (1314) continuously (e.g., first driving the solution containing encapsulated mRNA particles and then driving the solution containing beads; or first driving the solution containing beads and then driving the solution containing encapsulated mRNA particles). In the pattern of continuously driving different solutions towards the first pump (1314), one valve (1310, 1316) can be kept in the closed state while the other valve (1310, 1316) is open.

[0206] Once both solutions reach the first pump (1314), the two valves (1310, 1316) can be closed; and the first pump (1314) can be activated to drive both solutions towards the DLS chamber (1324), as shown by the box (1402). The third valve (1328) can be kept in the closed state during this part of the process. On the way to reaching the DLS chamber (1324), the solutions will pass through the third channel (1320) and the mixing chamber (1322) such that the DLS chamber (1324) will receive a mixture of the two solutions.

[0207] At this stage, the first pump (1314) can be deactivated and the valves (1310, 1316, 1328) can be kept closed such that the mixture remains in the DLS chamber (1324). With the mixture being held in the DLS chamber (1324), the Figures 9 to 11 and Figures 16 to 20 DLS components shown above can be activated to perform DLS on the mixture and thereby perform DLS measurements on the mixture, as shown by the box (1404). This DLS measurement can include projecting light into the mixture, receiving the light scattered by the beads and particles in the mixture, and tracking the change in the scattering pattern over time as the beads and particles disperse in the liquid by Brownian motion. The DLS data can be used to generate an autocorrelation curve, as shown by the box (1406) and also as described in more detail above.

[0208] Figure 27An illustration (1450) is shown that includes examples of autocorrelation curves (1452, 1454, 1456, 1458) that can be obtained via a DLS chamber (1324). For example, curve (1452) represents an autocorrelation curve generated from a solution containing encapsulated mRNA particles, where the encapsulated mRNA particles have a relatively small diameter (e.g., about 60 nm). Curve (1458) represents an autocorrelation curve generated from a solution containing beads, where the beads have a relatively large diameter (e.g., about 600 nm). Curve (1454) represents an autocorrelation curve generated from a mixture of a solution containing encapsulated mRNA particles and a solution containing beads. Curve (1456) represents an example of an autocorrelation curve corresponding to a specific single particle size population based on fitting the autocorrelation curve from a two-population solution to a single-population model. Curve (1456) represents that the autocorrelation curve generated from a solution containing two populations is different from the autocorrelation curve of a solution containing a monodisperse (single) population.

[0209] Returning to Figure 26 the process shown, using the data from the autocorrelation curve, the next step may include applying the data to the following equation (II) to determine the viscosity of the solution containing encapsulated mRNA particles, as shown in block (1408):

[0210] (II)

[0211]

[0212] where "y" is the autocorrelation value,

[0213] "A" is the vertical offset of the autocorrelation curve,

[0214] "B" is the magnitude of the exponent associated with a single population (e.g., encapsulated mRNA particles),

[0215] "Γ 1 " is the autocorrelation curve fitting value associated with the encapsulated mRNA particles, determined by fitting to the autocorrelation curve,

[0216] "D" is the magnitude of the exponent associated with another single population (e.g., beads),

[0217] "Γ 2 " is the autocorrelation curve fitting value associated with the beads, determined by fitting to the autocorrelation curve, and

[0218] "x" is the time lag of the autocorrelation.

[0219] The "Γ 1 " and "Γ 2The "Γ" values can each independently conform to Equation (I), where the larger "Γ" value corresponds to the larger known bead diameter. Thus, Equation (I) can be effectively rewritten as the following Equation (III):

[0220] (III)

[0221]

[0222] where "v" is the viscosity of the mixture of the aliquot and the bead solution,

[0223] “Γ 2 ” is the autocorrelation curve fitting value associated with the bead, determined by fitting to the autocorrelation curve,

[0224] “d 小珠 ” is the diameter of the bead (the value of which is known in this example), and

[0225] “K” is the remaining parameter in Equation (I), denoted as “K” in Equation (III) for simplicity.

[0226] Equation (III) can thus be solved to determine the viscosity of the mixture, as shown in block (1408). It should be understood that this will represent the viscosity of the combination of the solution containing the encapsulated mRNA particles and the solution containing the beads. It should also be understood that in this example, beads of known particle size are used as a calibration tool for determining the viscosity.

[0227] Once the viscosity of the mixture has been determined, the following Equation (IV) can be used to determine the particle size of the encapsulated mRNA particles:

[0228] (IV)

[0229]

[0230] where “d 颗粒 ” is the diameter of the encapsulated mRNA particles,

[0231] “Γ 1 ” is the autocorrelation curve fitting value associated with the encapsulated mRNA particles, determined by fitting to the autocorrelation curve,

[0232] “v” is the viscosity of the mixture (e.g., as determined via Equation (III)), and

[0233] “K” is the remaining parameter in Equation (I), denoted as “K” in Equation (III) for simplicity.

[0234] Once the mixture viscosity and the particle size of the encapsulated mRNA particles are determined based on the boxes (1408, 1410), the third valve (1328) can be transitioned to an open state, and the first pump (1324) can be activated to drive the mixture towards the second pump (1332) via the fourth channel (1326) and the fifth channel (1330). The fourth valve (1336) can be transitioned to an open state, and the second pump (1332) can be activated to drive the mixture towards the outlet channel (1306) via the sixth channel (1334). In some embodiments, the mixture is received in a waste storage compartment after leaving the measurement stage (1300) via the outlet channel (1306). In some other embodiments, a therapeutic composition is formed using the mixture after leaving the measurement stage (1300) via the outlet channel (1306). Alternatively, the mixture can be processed in any other suitable manner after leaving the measurement stage (1300) via the outlet channel (1306).

[0235] The process described above with reference to Figures 26 to 27 can be repeated to test different aliquots of the solution containing the encapsulated mRNA particles. In some embodiments, the valve (854) is operated to sequentially direct the output of each mixing assembly (820) towards the measurement stage (1300). The measured particle size and / or particle size distribution of the encapsulated mRNA particles can be compared to a threshold or a predetermined range to determine whether the actual particle size and / or particle size distribution of the encapsulated mRNA particles is within tolerance. This comparison can indicate whether the mixing assembly (820) that produced the aliquot processed by the measurement stage (1300) is operating properly. If the particle size and / or particle size distribution of the encapsulated mRNA particles is outside the tolerance, this can indicate that the particle size and / or particle size distribution of the encapsulated mRNA particles in the measurement stage (1300) is inappropriate; and the mixing assembly (820) that produced the aliquot in the measurement stage (1300) is operating improperly. The processing chip (800) can then stop the delivery of the output of that mixing assembly (820).

[0236] Regardless of whether the output of the mixing assembly (820) has produced an acceptable or unacceptable particle size and / or particle size distribution, after the output of the first mixing assembly (820) has been Figures 26 to 27After the process described above is analyzed in the measurement stage (1300), the processing chip (800) can operate the valve (854) to transfer the output of the next mixing component (820) to the measurement stage (1300). This sequence can continue until the output of each mixing component (820) has been analyzed in the measurement stage (1300). Once the output of the last mixing component (820) has been analyzed in the measurement stage (1300), the process can start from the first mixing component (820) and continue to execute this sequence throughout the entire duration of the operation of the processing chip (800). In the case where any mixing component (820) stops working due to the failure of its output in the measurement stage (1300), such mixing components (820) can be ignored as the processing chip (800) repeats the sequence of testing the output of the mixing components (820) in the measurement stage (1300).

[0237] In the measurement stage (1300), beads of known particle size are used as a calibration tool as described above with reference to Figures 26 to 27 In some variations of measuring the viscosity of the measurement solution described above, the second pump (1332) and the third valve (1328) or the fourth valve (1336) are omitted. In other words, the above process of using beads of known particle size as a calibration tool to measure the viscosity of the solution can be performed without the second pump (1332) and the third valve (1328) or the fourth valve (1336). The above process of using beads of known particle size as a calibration tool to measure the viscosity of the solution can also be performed using other variations of the measurement stage (1300), such that Figure 25 the features and arrangements shown are not necessarily the only features and arrangements available for performing the process.

[0238] In addition to the foregoing teachings specifically provided in the context of the measurement stage (1300) and Figures 26 to 27 the processing chip (800) including the measurement stage (1300) can be operated according to other teachings provided above regarding the operation of the processing chip (800).

[0239] Like other calculations and algorithms performed herein, Figure 26 the process shown can be executed by the controller (121). This can include the controller (121) activating the drive valves (1310, 1316, 1328, 1336), pumps (1314, 1332) and Figures 9 to 11 and Figures 16 to 20 the components of the DLS components shown. This can also include the controller (121) processing the autocorrelation data from the autocorrelator (1024) to generate an autocorrelation curve and performing the calculations associated with blocks (1408, 1410) as described above. Of course, the controller (121) can perform various other functions in combination with Figure 26 the process shown, including but not limited to other functions explicitly described herein.

[0240] C. Examples of Viscosity Sensing Using Serial Dilution

[0241] As a supplement or alternative to using beads of known particle size as a calibration tool to measure the viscosity of a solution, the viscosity of a solution can be measured by providing a controlled series of dilutions of the solution and tracking the changes in the solution based on the series of dilutions. An example of such a series of dilution processes is as Figure 28 shown. In this example, an aliquot of a solution containing encapsulated mRNA particles is conveyed towards the DLS chamber (1324), and an initial DLS measurement is obtained, as shown in the box (1500). This includes opening the first valve (1310) and driving an aliquot of the solution containing encapsulated mRNA particles towards the first pump (1314) via the channels (1302, 1312). Once the aliquot reaches the first pump (1314), the first valve (1310) is closed; then the first pump (1314) is activated to drive the aliquot via the channel (1320) and the mixing chamber (1322) into the DLS chamber (1324). The third valve (1328) can be kept closed during this part of the process. Alternatively, the third valve (1328) can be in an open state, and the aliquot can reside in the mixing chamber (1322), the DLS chamber (1324), and the second pump (1332). In the case where at least a portion of the aliquot is held in the DLS chamber (1324), the Figures 9 to 11 and Figures 16 to 20 DLS components shown above can be activated to perform DLS on the aliquot and thereby make an initial or baseline DLS measurement of the aliquot.

[0242] Once an initial DLS measurement of the aliquot has been made, a diluent is added to the aliquot as shown in block (1502). By way of example only, the diluent may include a buffer or any other suitable type of diluent. Additionally, the viscosity of the diluent may be known. This part of the process may include opening a second valve (1316) and driving a first volume of diluent towards a first pump (1314) via channels (1304, 1318). In some forms, the first volume of diluent exceeds the capacity of the first pump (1314) such that at least some of the first volume of diluent is conveyed through the first pump (1314) in the measurement stage (1300). Once the first volume of diluent has been introduced into the measurement stage (1300), the second valve (1316) is closed; then the first pump (1314) is activated to drive the first volume of diluent via channels (1320) and a mixing chamber (1322) into a DLS chamber (1324) such that the first volume of diluent is combined with an aliquot of the solution containing encapsulated mRNA particles in the DLS chamber (1324). In some forms, the combined volume of the first volume of diluent and the aliquot of the solution containing encapsulated mRNA particles is approximately equal to the combined capacity of the mixing chamber (1322), the DLS chamber (1324), at least one pump (1314, 1332), and channels (1320, 1326, 1330).

[0243] To further mix the first volume of diluent and the aliquot of the solution containing encapsulated mRNA particles, the pumps (1314, 1332) may be activated in sequence a predetermined number of repetitions. For example, valves (1310, 1316, 1336) may be kept in a closed state while valve (1328) is kept in an open state, and then the first pump (1314) may be activated to drive the mixture towards a second pump (1332). Next, the second pump (1332) may be activated to drive the mixture back towards the first pump (1314). In this way, the pumps (1314, 1332) may be activated alternately any suitable number of times. As the mixture flows between the pumps (1314, 1332), the mixture may repeatedly flow through the mixing chamber (1322), which may further facilitate mixing of the first volume of diluent with the aliquot of the solution containing encapsulated mRNA particles.

[0244] Once the first volume of diluent has been properly mixed with the aliquot of the solution containing encapsulated mRNA particles, even in scenarios where the pumps (1314, 1332) are not alternately activated to further mix the first volume of diluent with the aliquot of the solution containing encapsulated mRNA particles, the mixture may be held in the DLS chamber (1324); and may be activated as described above Figures 9 to 11 and Figures 16 to 20The DLS component shown performs DLS on the mixture and thereby makes a DLS measurement of the mixture, as shown in block (1504). The DLS measurement can include projecting light into the mixture, receiving the light scattered by beads and particles in the mixture, and tracking the change in the scattering pattern over time as the beads and particles disperse in the liquid by Brownian motion. The DLS data can be used to generate an autocorrelation curve, as shown in block (1506) and also as described in more detail above.

[0245] Thereafter, several additional volumes of diluent can be added to the mixture to further dilute the mixture. Each additional volume of diluent can have the same volume as the first volume of diluent. Each additional volume of diluent can also be added to (and mixed with) the previously formed mixture of diluent and aliquot according to the above procedure. Thus, part of the process includes determining whether an additional volume of diluent should be added, as shown in block (1508). In some embodiments, an additional volume of diluent (block (1502)) is added, a corresponding DLS measurement (block (1504)) is made, and a corresponding autocorrelation curve is fit (block (1506)) a predetermined number of times. In some other embodiments, the data obtained through the DLS measurement process is monitored to determine whether sufficient data has been obtained such that the number of iterations of dilution, DLS measurement, and autocorrelation curve fitting can vary with the process.

[0246] In embodiments using a predetermined number of iterations, the controller (121) or some other component can keep track of the number of times an additional volume of diluent has been added, as shown in block (1506); and can ensure that an additional volume of diluent has been added, a corresponding DLS measurement (block (1504)) has been made, and a corresponding autocorrelation curve has been fit (block (1506)) until a predetermined number of diluent volumes have been added. Similarly, the controller (121) can keep track of the data obtained through the DLS measurement process to determine whether sufficient data has been obtained; and can stop further iterations once the controller (121) determines that sufficient data has been obtained. In either case, the controller (121) or some other component can drive the subroutines for serial dilution, corresponding DLS measurement, and corresponding autocorrelation curve fitting. In some embodiments, each iteration of dilution (block (1502)) provides a 50% dilution. Alternatively, any other suitable dilution rate can be used, although it is still advantageous to provide the same dilution rate for each iteration of dilution.

[0247] Once the appropriate volume of diluent has been added (e.g., the dilution action (block (1502)) has been iterated a predetermined number of times or the data display indicates that no further iteration is required, etc.), the corresponding DLS measurement has been performed (block (1504)) and the corresponding autocorrelation curve has been fitted (block (1506)), the viscosity of the diluted mixture can be determined, as shown in block (1510). In this example, the encapsulated mRNA particles are delivered in a fluid medium containing ethanol such that the solution containing the encapsulated mRNA particles includes the encapsulated mRNA particles and ethanol; and the diluent includes water. Alternatively, any other suitable fluid medium can be used to deliver the encapsulated mRNA particles; and any other fluid can be used as the diluent. Returning to this example, although the exact amount of ethanol in the solution containing the encapsulated mRNA particles may be unknown, it can be assumed that the amount of ethanol in the solution containing the encapsulated mRNA particles is approximately a certain amount (e.g., about 15%, etc.). The viscosity of the mixture can increase linearly with the ethanol concentration, as shown in the following equation (V):

[0248] (V)

[0249] v = f([ethanol]) ≈ m[ethanol] + b

[0250] where "v" is the viscosity of the mixture of the aliquot and the diluent,

[0251] "f([ethanol])" is the viscosity as a linear function of the ethanol content,

[0252] "m([ethanol])" is the linearized term in the equation, representing the concentration of ethanol, and

[0253] "b" is the linearized term in the equation, representing the initial viscosity without any ethanol.

[0254] In the case of determining the viscosity of the mixture using equation (V) as described above, the next step of the process involves solving for the particle size of the encapsulated mRNA particles, as shown in block (1512). To do this, since the concentration of ethanol in the mixture decreases by a known factor ("f") with each iteration of dilution (block (1502)), the value of the autocorrelation gamma value ("Γ") relative to the number of dilution iterations can be determined according to the following equation (VI):

[0255] (VI)

[0256]

[0257] where "Γ i " is the autocorrelation curve fitting value determined by fitting the autocorrelation curve for each dilution iteration (block (1502)),

[0258] "K" is the remaining parameter in equation (I), denoted as "K" in equation (III) for simplicity,

[0259] “d” is the diameter of the encapsulated mRNA particles,

[0260] “b” is the linearized term in the equation, representing the initial viscosity without any ethanol,

[0261] “m([ethanol]) 0 ” is the linearized term in the equation, representing the initial concentration of ethanol,

[0262] “f i ” is the dilution factor determined by the microfluidic volume (e.g., if the input microfluidic buffer dilutes the sample by half each time, then each time it will have a dilution factor of 50%, and this will be determined by the buffer volume introduced for each dilution cycle, as determined by the microfluidic geometry / volume),

[0263] “A” is the fitting parameter, representing K / d,

[0264] “B” is the fitting parameter, representing b, and

[0265] “C” is the fitting parameter, representing m([ethanol]) 0 .

[0266] In this example, the value of “A” enables determination of the particle size of the encapsulated mRNA particles in the aliquot; and the value of “C” enables determination of the concentration of ethanol in the aliquot.

[0267] Figure 29 A diagram (1550) is shown that includes an example of a curve (1552), plotting an example of the value of “Γ i ” during 20 dilution iterations (box (1502)), where each dilution iteration (box (1502)) provides a 50% dilution rate. As shown, the value of “Γ i ” increases significantly from approximately the third dilution (box (1502)) iteration to approximately the ninth dilution iteration; and then remains essentially flat starting from approximately the tenth dilution (box (1502)) iteration. Thus, in this example, it can be determined that the dilution (box (1502)) can be iterated between approximately three and nine times, and additional dilution (box (1502)) iterations may not necessarily be beneficial. It should be understood that this is merely an illustrative example, and different processes may require more or fewer dilution iterations (box (1502)).

[0268] According to the foregoing, with each repetition of the dilution step (box (1502)), the sample concentration in the measurement stage (1300) decreases by a known factor specific to the design of the measurement stage (1300). This factor can be proportional to the volumes of the pumps (1314, 1332), mixing chamber (1322), DLS chamber (1324), etc.

[0269] The process described above with reference can be repeated Figures 28 to 29 to test different aliquots of the solution containing the encapsulated mRNA particles. In some embodiments, the valve (854) is operated to sequentially direct the output of each mixing assembly (820) towards the measurement stage (1300). The measured particle size and / or particle size distribution of the encapsulated mRNA particles can be compared to a threshold or a predetermined range to determine whether the actual particle size and / or particle size distribution of the encapsulated mRNA particles is within tolerance. This comparison can indicate whether the mixing assembly (820) that produced the aliquot processed by the measurement stage (1300) is operating properly. If the particle size and / or particle size distribution of the encapsulated mRNA particles is outside the tolerance, this can indicate that the particle size and / or particle size distribution of the encapsulated mRNA particles in the measurement stage (1300) is inappropriate; and the mixing assembly (820) that produced the aliquot in the measurement stage (1300) is operating improperly. The controller (121) can then effectively shut down the mixing assembly (820), as described herein.

[0270] Regardless of whether the output of the mixing assembly (820) has produced an acceptable or unacceptable particle size and / or particle size distribution, after the output of the first mixing assembly (820) has been analyzed in the measurement stage (1300) according to the process described above with reference Figures 28 to 29 the controller (121) can operate the processing chip (800) by actuating the valve (854) to transfer the output of the next mixing assembly (820) to the measurement stage (1300). This sequence can continue until the output of each mixing assembly (820) has been analyzed in the measurement stage (1300). Once the output of the last mixing assembly (820) has been analyzed in the measurement stage (1300), the process can start from the first mixing assembly (820) and continue to execute this sequence for the entire duration of the operation of the processing chip (800). In the event that any mixing assembly (820) stops working due to a failed output in the measurement stage (1300), such mixing assemblies (820) can be ignored as the processing chip (800) repeats the sequence of testing the output of the mixing assemblies (820) in the measurement stage (1300).

[0271] In addition to the foregoing teachings specifically provided in the context of the measurement stage (1300) and Figures 28 to 29 the processing chip (800) including the measurement stage (1300) can be operated according to other teachings provided above regarding the operation of the processing chip (800).

[0272] Like other calculations and algorithms performed herein, Figure 28The process shown can be performed by a controller (121). This can include the controller (121) activating drive valves (1310, 1316, 1328, 1336), pumps (1314, 1332) and Figures 9 to 11 and Figures 16 to 20 components of the DLS components shown. This can also include the controller (121) processing autocorrelation data from the autocorrelator (1024) to generate an autocorrelation curve and performing the calculations associated with blocks (1408, 1410) as described above. Of course, the controller (121) can perform various other functions in conjunction with Figure 28 the process shown, including but not limited to other functions explicitly described herein.

[0273] V. Examples of Processing Chips with Fluid Input Manifolds

[0274] As described above, the processing chip (800) can include a set of channels (856, 858, 860) that cooperate to form an output manifold. This allows the outputs of several mixing components (820) to be delivered to a single location, such as the measurement stage (1300). In some cases, it may be desirable to provide an input manifold that allows fluid from a single input source to reach the corresponding input channels of several mixing components (820). This can avoid the need to have a dedicated set of fluid input ports (804a, 804b, 804c) for each mixing component (820). This can in turn reduce the manufacturing and operational complexity associated with the processing chip. Figures 30 to 33 An example of a processing chip (2000) that can provide such benefits is shown.

[0275] The processing chip (2000) of this example includes three fluid input ports (2004a, 2004b, 2004c) and three corresponding fluid channels (2002a, 2002b, 2002c). By way of example only, in some specific implementations, a combination of mRNA and water is delivered through fluid input port (2004a) and fluid channel (2002a), one or more delivery carrier molecules in ethanol are delivered through fluid input port (2004b) and fluid channel (2002b), and a diluent is delivered through fluid input port (2004c) and fluid channel (2002c). Alternatively, any other suitable fluid can be delivered through fluid input ports (2004a, 2004b, 2004c) and fluid channels (2002a, 2002b, 2002c). In this example, fluid channel (2002a) leads to fluid input manifold channel (2005a), which further leads to a plurality of mixing components (2020), such that each mixing component (2020) can receive fluid from fluid input port (2004a) via channels (2002a, 2005a). Fluid channel (2002b) leads to fluid input manifold channel (2005b), which further leads to a plurality of mixing components (2020), such that each mixing component (2020) can receive fluid from fluid input port (2004b) via channels (2002b, 2005b). Fluid channel (2002c) leads to fluid input manifold channel (2005c), which further leads to a plurality of mixing components (2020), such that each mixing component (2020) can receive fluid from fluid input port (2004c) via channels (2002c, 2005c).

[0276] As Figures 30 to 31 shown, the processing chip (2000) of this example includes three pressure sensing regions (2010). The first pressure sensing region (2010a) is fluidly coupled to the first fluid channel (2002a), such that the first pressure sensing region (2010a) can detect the pressure of the fluid within the first fluid channel (2002a). The second pressure sensing region (2010b) is fluidly coupled to the second fluid channel (2002b), such that the second pressure sensing region (2010b) can detect the pressure of the fluid within the second fluid channel (2002b). The third pressure sensing region (2010c) is fluidly coupled to the third fluid channel (2002c), such that the third pressure sensing region (2010c) can detect the pressure of the fluid within the third fluid channel (2002c). The pressure sensing regions (2010) can be part of a pressure sensing stage configured and operable like the pressure sensing stage (700) described above. The pressure sensing regions (2010) can also be configured and operable like the pressure sensing stage (810) described above. InFigure 31 In the case where certain features are omitted, it should be understood that Figure 31 only the pressure sensing chambers (2012) of the pressure sensing area (2010) are shown.

[0277] Figure 32 The features of the mixing assembly (2020) are shown in more detail. Although only one mixing assembly (2020) is shown in Figure 32 , it should be understood that all the mixing assemblies (2020) in the processing chip (2000) can be configured and operable like the Figure 32 shown mixing assembly (2020). As shown, each mixing assembly (2020) includes a set of vacuum covers (2022), a set of inlet valves (2024), and a set of mixing chambers (2030, 2040). The first vacuum cover (2022a) receives fluid from the first fluid channel (2002a) via the fluid input manifold channel (2005a). The second vacuum cover (2022b) receives fluid from the second fluid channel (2002b) via the fluid input manifold channel (2005b). The third vacuum cover (2022c) receives fluid from the third fluid channel (2002c) via the fluid input manifold channel (2005c). The first valve (2024a) meters the flow rate of the fluid from the first vacuum cover (2022a) to the first channel (2026a) leading to the first mixing chamber (2030). The second valve (2024b) meters the flow rate of the fluid from the second vacuum cover (2022b) to the inlet channel (2026b) leading to the first mixing chamber (2030). The channels (2026a, 2026b) converge to form an inlet channel (2032) leading to the first mixing chamber (2030). The fluids from the channels (2026a, 2026b) are thus mixed together within the first mixing chamber (2030).

[0278] The third valve (2024c) meters the flow rate of the fluid from the third vacuum cover (2022c) to the third channel (2026c) leading to the second mixing chamber (2040). The outlet channel (2034) from the first mixing chamber (2030) converges with the third channel (2026c) to form an inlet channel (2042) leading to the second mixing chamber (2040). The fluids from the channels (2034, 2026c) are thus mixed together within the second mixing chamber (2040). The fluid mixed in the second mixing chamber (2040) is output via the outlet channel (2044).

[0279] In some forms of using a mixing assembly to provide encapsulated mRNA, a combination of mRNA and water can be delivered through a first fluid channel (2002a), and one or more delivery carrier molecules in ethanol can be delivered through a second fluid channel (2002b). In this type of form, the mRNA and the delivery carrier molecules can thus be combined in a first mixing chamber (2030) for encapsulation. A diluent (e.g., a citrate-based buffer solution, etc.) can be delivered through a third fluid channel (2002c). In this type of form, a second mixing chamber (2040) can thus be used to provide pH adjustment. In some variations, the mRNA and water are combined in another mixing chamber (not shown) upstream of the first fluid channel (2002a). Similarly, the delivery carrier molecules and ethanol can be combined in another mixing chamber (not shown) upstream of the second fluid channel (2002b).

[0280] A valve (2054) is positioned downstream of the outlet channel (2044); a common manifold channel (2058) is positioned downstream of the valve (2054). All mixing assemblies (2020) are fluidly coupled to the common manifold channel (2058) in such a way that the valve (2054) is interposed between the respective mixing assembly (2020) and the common manifold channel (2058). If the valve (2054) associated with a particular mixing assembly (2020) is in a closed state, fluid cannot be transferred from that mixing assembly (2020) to the common manifold channel (2058). If the valve associated with a particular mixing assembly (2020) is in an open state, fluid can be transferred from that mixing assembly (2020) to the common manifold channel (2058). Thus, the valve (2054) can be selectively opened and closed to selectively provide fluid communication between the selected mixing assembly (2020) and the common manifold channel (2058).

[0281] Insofar as the drawings may appear to imply that the valve (2054) is positioned along the manifold channel (2058), in this example, the valve (2054) is not actually positioned to prevent fluid flow along the manifold channel (2058). Instead, each valve (2054) is only positioned to prevent fluid from its corresponding mixing assembly 2020 from flowing into the manifold channel (2058). In this example, no valve (2054) will affect the flow of fluid from any other mixing assembly (2020); any valve (2054) will also not prevent fluid output from another mixing assembly 2020 from flowing through the manifold channel (2058). Thus, each valve (2054) will only prevent fluid from its corresponding mixing assembly (2020) from flowing into the manifold channel (2058), without preventing fluid from any other mixing assembly (2020) from flowing along the manifold channel (2058).

[0282] In this example, the controller (121) is configured to drive the valves (2054) in such a way that only one valve (2054) is in the open state at a given moment of operation. At some stages of operation, all the valves (2054) may be in the closed state. At some other stages of operation, two or more valves (2054) may be in the open state. The features of the processing chip (2000) downstream of the common manifold channel (2058) will be described in more detail below.

[0283] As described above, the processing chip (2000) may operate in such a way that based on the operating state of each valve (2054), the fluid outputs of the mixing assemblies (2020) can all be delivered to the same common manifold channel (2058). In other words, the fluid outputs of the mixing assemblies (2020) can ultimately reach the same destination, the common manifold channel (2058). Also as described above, the mixing assemblies (2020) may all receive fluid from the same input source, namely a fluid source coupled to the fluid input ports (2004a, 2004b, 2004c). This is due to the existence of corresponding fluid input manifold channels (2005a, 2005b, 2005c) shared by the mixing assemblies (2020).

[0284] Just as the controller (121) can be configured to drive the valve (2054) in such a way that at a given operating moment, only the fluid output of one mixing component (2020) can be transmitted to the same common manifold channel (2058), the controller (121) can be configured to allow only one mixing component (2020) to receive fluid from the fluid input ports (2004a, 2004b, 2004c) at a given operating moment. For example, the processing chip (200) can operate in such a way that at a given operating moment, only the inlet valve (2024) of one mixing component (2020) is in the open state while the inlet valves (2024) of all other mixing components (2020) are in the closed state. The valve (2024) can thus be activated to provide fluid communication from the fluid input ports (2004a, 2004b, 2004c) to only one selected mixing component (2020) at a given operating moment. In some stages of operation, the valves (2024) of all the mixing components 2020 can be in the closed state. In some other stages of operation, the valves (2024) of two or more mixing components (2020) can be in the open state. Examples of scenarios in which the controller (121) can activate the valve (2024) to effectively switch from one mixing component (2020) to another will be described in more detail below. Although the valve (2024) is described herein as providing a structure for selectively closing the fluid communication with the mixing component (2020), some other forms can include a valve with similar operability that is positioned upstream of the vacuum cover (2022) (i.e., between the fluid input manifold channels (2005a, 2005b, 2005c) and the corresponding vacuum covers (2022a, 2022b, 2022c)).

[0285] Figure 33More particularly, features of the processing chip (2000) downstream of the common manifold channel (2058) (and thus, downstream of the mixing assembly (2020)) are shown. As shown, the common manifold channel (2058) leads to a manifold outlet channel (2060). The manifold outlet channel (2060) is fluidly coupled to a set of valves (2062, 2064, 2066). The valve (2064) is fluidly coupled to a waste channel (2200), which is fluidly coupled to a waste output port (2202). The waste output port (2202) can be used to transfer fluid to a waste storage compartment. Such a waste storage compartment can be located in another processing chip (111, 200, 500, 800, 2000), in a reagent storage rack (107), or elsewhere. Examples of scenarios in which fluid can be transferred through the waste output port (2202) will be described in more detail below. It should be understood that the valves (2062, 2064) can be selectively transitioned between a closed state and an open state to allow fluid from the manifold outlet channel (2060) to be transferred to the waste output port (2202) via the waste channel (2200).

[0286] The valve (2066) is fluidly coupled to an output channel (2100), which is fluidly coupled to a main output port (2102). In some configurations, the fluid from the main output port (2102) constitutes an acceptable therapeutic composition. In some other configurations, the fluid from the main output port (2102) is further processed to form a therapeutic composition. Such further processing can include transferring the fluid to another processing chip (111, 200, 500, 800, 2000), to a vial in a reagent storage rack (107), or elsewhere. As another alternative, the fluid from the main output port (2102) can be used for some other quality control tests (e.g., on another processing chip (111, 200, 500, 800, 2000) or elsewhere). Alternatively, the fluid from the main output port (2102) can be processed in any other suitable manner. It should be understood that the valves (2062, 2066) can be selectively transitioned between a closed state and an open state to allow fluid from the manifold outlet channel (2060) to be transferred to the main output port (2102) via the output channel (2100).

[0287] In addition to being fluidly coupled to the manifold outlet passage (2060), the manifold passage (2058) is also fluidly coupled to the measurement stage (2300). The measurement stage (2300) of this example is configured and operates similar to the measurement stage (1300) described above. The measurement stage (2300) of this example includes a sample inlet passage (2302), a test fluid inlet passage (2400), and an outlet passage (2306). A valve (2301) is interposed between the sample inlet passage (2302) and the manifold passage (2058) such that the valve (2301) can be selectively opened to provide an aliquot (i.e., a sample volume) of the solution carrying encapsulated mRNA particles output from the mixing assembly (2020) to the sample inlet passage (2302). When the valve (2301) is in the open state to provide an aliquot to the measurement stage (2300) via the sample inlet passage (2302), the valve (2062) can be in the closed state.

[0288] The sample inlet passage (2302) is also fluidly coupled to a passage (2506) having an associated valve (2504). The valve (2301) is positioned at the junction between the sample inlet passage (2302), the passage (2506), and the manifold passage (2058). Another passage (2500) is also fluidly coupled to the valve (2504). The passage (2500) leads to a port (2502), as Figures 30 to 31As shown. In this example, port (2502) is used to transfer a flushing fluid to processing chip (2000). By way of example only, the flushing fluid can include a combination of water and ethanol and / or any other suitable type of flushing fluid. Valve (2504) can selectively transition between a closed state and an open state to prevent or allow fluid communication from port (2502) and channel (2500) to channel (2506). In some configurations, when the flushing fluid is transferred through channel (2500) via port (2502), valves (2504, 2301, 2310, 2336) are in an open state, while valves (2054, 2316, 2064, 2066) are in a closed state, such that the flushing fluid passes through measurement stage (2300) and ultimately exits processing chip (2000) via waste output port (2202). Alternatively, the various valves within processing chip (2000) can be selectively opened or closed in any other suitable scheme to provide any other desired flow path of the flushing fluid through processing chip (2000). By way of example only, when the flushing fluid is transferred through channel (2500) via port (2502), at least valves (2066, 2310, 2336) can be closed, and valves (2301, 2054, 2064) can be opened, such that the flushing fluid passes through common manifold channel (2058) and ultimately exits processing chip (2000) via waste output port (2202). In some other configurations, channels (2500, 2506), port (2502), and valve (2504) are omitted.

[0289] Test fluid inlet channel (2400) is in fluid communication with a test fluid source (not shown) via test fluid port (2402). As described above in the context of measurement stage (1300), the test fluid can include a fluid with beads, a diluent without beads (e.g., buffer and water, etc.), and / or any other suitable type of fluid. Outlet channel (2306) can be considered similar to outlet channel (1306) such that a solution can exit measurement stage (2300) via outlet channel (2306). As Figure 33 shown, outlet channel (2306) ultimately leads to waste output port (2202) via waste channel (2200). During the operation stage where fluid is transferred from measurement stage (2300) to waste output port (2202) via waste channel (2200), valve (2064) can be in a closed state to prevent such fluid from reaching manifold outlet channel (2060).

[0290] The metering stage (2300) of this example also includes a first valve (2310), a first channel (2312), a first pump (2314), a second valve (2316), a second channel (2318), a third channel (2320), a mixing chamber (2322), a DLS chamber (2324), a fourth channel (2326), a second pump (2332), a fifth channel (2334), and a third valve (2336). The valves (2310, 2316, 2336) of this example can be configured and operable like the above-described valves (824, 854). Thus, the valves (2310, 2316, 2336) can drive the elastic layer (e.g., like the elastic layer (302)) of the processing chip (2000) to transition between an open state and a closed state by selectively applying pressure (or releasing pressure on the elastic layer) to the elastic layer in the area of the processing chip (2000) corresponding to the valves (2310, 2316, 2336). The valves (2310, 2316, 2336) can thus be used to selectively block or allow fluid flow. The other valves (2024, 2054, 2064, 2066, 2301, 2504) of the processing chip (2000) can be configured and operated in a similar manner. The pumps (2314, 2332) can also be operated by selectively applying pressure to the elastic layer (e.g., like the elastic layer (302)) of the processing chip (2000) in the area of the processing chip (2000) corresponding to the pumps (2314, 2332), thereby driving fluid flow through a peristaltic pumping action.

[0291] The first valve (2310) is interposed between the sample inlet channel (2302) and the first channel (2312) such that the first valve (2310) can be operated to selectively block or allow fluid to flow between the sample inlet channel (2302) and the first channel (2312). The first channel (2312) is also in fluid communication with the first pump (2314). The second valve (2316) is interposed between the test fluid inlet channel (2400) and the second channel (2318) such that the second valve (2316) can be operated to selectively block or allow fluid to flow between the test fluid inlet channel (2400) and the second channel (2318). The second channel (2318) is also in fluid communication with the first pump (2314). The first pump (2314) is also in fluid communication with the third channel (2320). The first pump (2314) can be operated to pump fluid from the sample inlet channel (2302) and / or the test fluid inlet channel (2400) towards the third channel (2320), depending on the state of each respective valve (2310, 2316).

[0292] The third channel (2320) is also in fluid communication with a mixing chamber (2322). The mixing chamber (2322) is configured to mix fluids from the sample inlet channel (2302) and the test fluid inlet channel (2304), such as being pumped into the mixing chamber (2322) via the first pump (2314). By way of example only, the mixing chamber (2322) can be configured and operable in the same manner as any mixing chamber described herein. Alternatively, the mixing chamber (2322) can be configured and operable in any other suitable manner.

[0293] The mixing chamber (2322) leads directly to the DLS chamber (2324). The DLS chamber (2324) can be constructed and operated in the same manner as the above-described DLS chamber (870). Thus, the body (900) can be positioned adjacent to the DLS chamber (2324) such that the source optical fiber (1010) can emit light into the DLS chamber (2324); and such that the sensing optical fiber (1020) can receive light scattered from particles in the fluid in the DLS chamber (2324) as described above. The scattered light can reach the photon counter (1022), and the corresponding data can be processed by the autocorrelator (1024), as described above.

[0294] The DLS chamber (1324) is also in fluid communication with a fourth channel (2326) leading to a second pump (2332). The second pump (2332) is also in fluid communication with a fifth channel (2334). The second pump (2332) is operable to pump fluid from the fourth channel (1326) towards the fifth channel (2334); and vice versa. In some configurations, another valve (not shown) can be interposed in the fluid path between the fourth channel (2326) and the second pump (2332). Such a valve can be configured and operable in the same manner as the third valve (1328) described above in the context of the measurement stage (1300). Returning to this example, the fifth channel (2334) is further in fluid communication with a third valve (2336). The third valve (2336) is also in fluid communication with the outlet channel (2306) such that the third valve (2336) is operable to selectively block or allow fluid flow between the fifth channel (2334) and the outlet channel (2306).

[0295] From the foregoing, it should be understood that the pumps (2314, 2332) can drive fluid towards and away from the DLS chamber (2324) from two separate sources. Additionally, the valves (2310, 2316, 2336) can selectively block fluid flow at different respective regions of the processing chip (2000) in conjunction with the operation of the pumps (2314, 2332). The measurement stage (2300) can be used in combination with the DLS components shown in Figures 9 to 11 and Figures 16 to 20 such that, as described above in the measurement stage (1300) and in conjunction with Figures 25 to 29Sense viscosity, particle size, and particle size density in the manner described in the context of the corresponding disclosure content.

[0296] As described above, at any given operating level, valve (204) can be selectively activated to provide fluid communication through only one mixing component (2020). In other words, valve (204) can be used to effectively designate one mixing component (2020) as "active" while effectively designating other mixing components (2020) as "inactive". Also as described above, valves (2062, 2064, 2066, 2301) can be selectively activated to determine whether the fluid output by the active mixing component (2020) will be conveyed to the measurement stage (2300), the main output port (2102), or the waste output port (2202). Regardless of where the fluid output by the active mixing component (2020) is conveyed, valves (204, 2054) of the inactive mixing components (2020) can remain closed.

[0297] Also as described above, the pressure sensing area (2010) and the measurement stage (2300) can be used to collect data related to the performance of the biochip (2000), including the performance specific to the currently active mixing component (2020). In terms of the controller (120) being able to operate to drive the operation of various valves (2024, 2054, 2062, 2064, 2066, 2301, 2504, 2310, 2316, 2336) of the processing chip (2000), and in terms of the controller (120) being able to operate to process data from the pressure sensing area (2010) and the measurement stage (2300), the controller (120) can be further configured to execute a control algorithm based on real-time feedback. In other words, the controller (120) can automatically drive valves (2024, 2054, 2062, 2064, 2066, 2301, 2504, 2310, 2316, 2336) and / or other features of the system (100) at least partially based on the real-time data collected via the pressure sensing area (2010) and the measurement stage (2300). In some cases, such automated operation via the controller (120) can include automated switching from one mixing component (2020) designated as active to another mixing component (2020) designated as active; the previously active mixing component (2020) becomes inactive.

[0298] In one example of operation, a first mixing component (2020) on a processing chip (2000) can be considered active, while other mixing components (2020) on the processing chip (2000) can be considered inactive. Valves (2064, 2301) can remain closed, and valves (2062, 2066) can remain open to direct fluid output from the first mixing component (2020) to the main output port (2102). Eventually, fluid can be temporarily redirected from the main output port (2102) to the measurement stage (2300) to test an aliquot of the fluid. This can be accomplished by closing valve (2062) and opening valve (2301), or performing the operations described above in the context of the measurement stage (2300). In some cases, such testing is performed periodically based on the passage of time. In some other cases, the test is performed in response to one or more other conditions (e.g., pressure data from one or more pressure sensing areas in the pressure sensing area (2010) falling outside of tolerance, etc.). Regardless of the basis for performing the aliquot test, the test process can be automatically initiated by the controller (120).

[0299] If the aliquot passes the test of the measurement stage (2300) (e.g., indicating that the particle size and / or distribution is within tolerance), the controller (120) can transition the processing chip (2000) back to the non-test mode, in which fluid output from the first mixing component (2020) is directed back to the main output port (2102). Valves (2064, 2301) can thus automatically return to the closed state; valves (2062, 2066) return to the open state. The aliquot processed by the measurement stage (2300) can leave the processing chip (2000) via the waste output port (2202).

[0300] The above process can be repeated until the controller (120) automatically determines that one or more conditions are outside the tolerance. In some cases, this may occur after a certain period of time when a given mixing component (2020) has been considered active. Additionally, or in an alternative form, the controller (120) can automatically determine that the particle size and / or distribution in the fluid output from the active mixing component (2020) is outside the tolerance based on data from the measurement stage (2300). Additionally, or in an alternative form, the controller (120) can automatically determine that the pressure of the fluid being delivered to the active mixing component (2020) is outside the tolerance based on data from one or more pressure sensing regions in the pressure sensing region (2010). Additionally, or in an alternative form, according to the above teachings, the controller 120 can automatically determine that the flow rate of the fluid being delivered through the active mixing component 2020 is outside the tolerance. Additionally, or in an alternative form, the controller (120) can automatically determine that any other condition associated with the active mixing component (2020) is outside the tolerance.

[0301] Once the controller (120) automatically determines that one or more conditions associated with the active mixing component (2020) are outside the tolerance. The controller (120) can automatically switch from one mixing component (2020) to another mixing component (2020). For example, the controller (120) can transition the valve (2024) of the first mixing component (2020) to the closed state to prevent fluid from reaching the first mixing component (2020) via the input manifold channel (2005); and transition the switching valve (2054) of the first mixing component (2020) to the closed state to prevent fluid from flowing back into the first mixing component (2020) via the common manifold channel (2058). Such closing of the valves (2024, 2054) can render the first mixing component (2020) inactive. The controller (120) can open the valves (2024, 2054) of the second mixing component (2020) to the open state. Such opening of the valves (2024, 2054) of the second mixing component (2020) can render the second mixing component (2020) active.

[0302] At this operating level, the routine of delivering the fluid output of the second mixing component (2020) to the main output port (2102) described above can be performed, and aliquots can be tested via the measurement stage (2300) until the controller (120) automatically determines that one or more conditions associated with the second mixing component (2020) are outside of tolerance. The controller (120) can thus cycle through the available mixing components (2020) on the processing chip (2000) until no mixing components (2020) remain. If the controller (120) determines that one or more conditions associated with the last remaining mixing component (2020) are outside of tolerance, the controller (120) can terminate further fluid processing on the processing chip (2000) and provide an appropriate notification to the operator.

[0303] By having only three fluid input ports (2004) and three corresponding fluid channels (2002) to provide a fluid communication path for all mixing components (2020), the processing chip (2000) can facilitate switching from one mixing component (2020) to another during the fluid mixing process. In the case where the mixing components (2020) share an input channel (2002) via a manifold channel (2005), it is not necessary to move and reconnect the fluid conduits from the reagent rack (107) from one set of input ports leading to one mixing component to another set of input ports leading to another mixing component. In the present example, switching between mixing components is simplified by using valves (2024, 2054).

[0304] VI. Examples of Combinations

[0305] The following examples relate to various non-exhaustive ways in which the teachings herein can be combined or applied. The following embodiments are not intended to limit the scope of any claims that may be presented at any time in the present application or a subsequent application of the present application. There is no disclaimer. The following embodiments are provided for illustrative purposes only. It is contemplated that the various teachings herein can be arranged and applied in many other ways. It is also contemplated that some variations may omit certain features mentioned in the following examples. Accordingly, unless explicitly stated by the inventors or successors-in-interest of the inventors at a later date, the aspects or features mentioned below should not be considered critical. If any claims are presented in the present application or a subsequent application related to the present application that include additional features other than those mentioned below, those additional features should not be considered to have been added for any reason related to patentability.

[0306] Example 1

[0307] An apparatus includes: a processing chip, the processing chip including: a first outer surface, a second outer surface, a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet, a light-transmissive material positioned between the first outer surface and the fluid chamber, a plurality of mixing components, each of the plurality of mixing components having a plurality of inlets and an outlet, each of the plurality of mixing components being configured to form a mixture of fluids from the plurality of inlets and convey the mixture through the outlet, and a fluid input manifold channel, at least one inlet of each of the plurality of mixing components being fluidly coupled to the fluid input manifold channel; and a dynamic light scattering component, the processing chip being removably positioned relative to the dynamic light scattering component, the dynamic light scattering component including: a body, the body including a first port and a second port, the body being positioned adjacent to the first outer surface, a first optical fiber coupled to the first port of the body, the first optical fiber for emitting light, the first port for guiding the light emitted by the first optical fiber through the light-transmissive material and into the fluid chamber, and a second optical fiber coupled to the second port of the body, the second optical fiber at the second port being inclined relative to the first optical fiber at the first port, the second optical fiber receiving light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber.

[0308] Example 2

[0309] The apparatus according to Embodiment 1, wherein the fluid chamber has a cylindrical shape including a circular upper inner surface, a circular lower inner surface, and an inner sidewall extending from the circular upper inner surface to the circular lower inner surface.

[0310] Example 3

[0311] The apparatus according to Embodiment 2, wherein the fluid chamber inlet is positioned in a region of the inner sidewall adjacent to the circular lower inner surface.

[0312] Example 4

[0313] The apparatus according to any one of Embodiments 2 to 3, wherein the fluid chamber outlet is positioned in a region of the inner sidewall adjacent to the circular upper inner surface.

[0314] Example 5

[0315] The apparatus according to any one of Embodiments 1 to 4, wherein at least one of the plurality of mixing components includes a first mixing stage for mixing a first plurality of fluid components to form a first fluid mixture, and the fluid chamber inlet is for receiving the first fluid mixture.

[0316] Example 6

[0317] The device according to Embodiment 5, the first mixing stage includes a first mixing inlet, a second mixing inlet, and a first mixing outlet. The first mixing inlet is for receiving a first fluid component, the second mixing inlet is for receiving a second fluid component, and the first mixing outlet is for outputting a first fluid mixture, which contains at least the first fluid component and the second fluid component.

[0318] Example 7

[0319] The device according to Embodiment 6, the processing chip further includes: a first pressure sensor for sensing the pressure of the first fluid component entering the first mixing inlet, and a second pressure sensor for sensing the pressure of the second fluid component entering the second mixing inlet.

[0320] Example 8

[0321] The device according to Embodiment 7, further includes a processor for receiving data from the dynamic light scattering component, the first pressure sensor, and the second pressure sensor.

[0322] Example 9

[0323] The device according to Embodiment 8, the processor is for further correlating the data received from the dynamic light scattering component, the first pressure sensor, and the second pressure sensor.

[0324] Example 10

[0325] The device according to any one of Embodiments 6 to 9, the processing chip further includes an additional fluid channel fluidly coupled to the first mixing outlet.

[0326] Example 11

[0327] The device according to Embodiment 10, the processing chip is for providing the transfer of fluid from the first mixing outlet to the additional fluid channel, the fluid chamber inlet, or a combination of the additional fluid channel and the fluid chamber inlet.

[0328] Example 12

[0329] The device according to any one of Embodiments 10 to 11, the processing chip is for providing the transfer of fluid from the additional fluid channel to the fluid chamber inlet via the first mixing outlet.

[0330] Example 13

[0331] The device according to any one of Embodiments 5 to 12, at least one of the plurality of mixing components further includes a second mixing stage having a second mixing outlet for mixing a second plurality of fluid components to form a second fluid mixture, and a fluid chamber inlet for receiving the second fluid mixture from the second mixing outlet.

[0332] Example 14

[0333] The device according to Embodiment 13, the processing chip further includes at least one valve for regulating the flow rate of the fluid from the first mixing outlet and the second mixing outlet to the fluid chamber inlet such that the fluid chamber inlet selectively receives only one of the first fluid mixture or the second fluid mixture at a time.

[0334] Example 15

[0335] The device according to any one of Embodiments 13 to 14, the processing chip further includes a manifold for guiding the fluid from the first mixing outlet and the second mixing outlet to the fluid chamber inlet.

[0336] Example 16

[0337] The device according to any one of Embodiments 1 to 15, the processing chip has a square shape including four corners, and the dynamic light scattering component is positioned at one of the four corners.

[0338] Example 17

[0339] The device according to any one of Embodiments 1 to 16, the dynamic light scattering component further includes a collimator located in the first port, and the collimator is inserted between the end of the first optical fiber and the first outer surface.

[0340] Example 18

[0341] The device according to Embodiment 17, the first port further includes a focusing volume inserted between the collimator and the first outer surface.

[0342] Example 19

[0343] The device according to Embodiment 18, the focusing volume defines a conical shape.

[0344] Example 20

[0345] The device according to any one of Embodiments 18 to 19, the dynamic light scattering component further includes a focusing lens inserted between the collimator and the focusing volume.

[0346] Example 21

[0347] For the device according to any one of Embodiments 1 to 20, the dynamic light scattering assembly further includes a filter located in the second port, and the filter is inserted between the end of the second optical fiber and the first outer surface.

[0348] Example 22

[0349] For the device according to Embodiment 21, the body further defines a channel inserted between the filter and the first outer surface.

[0350] Example 23

[0351] For the device according to any one of Embodiments 1 to 22, the body includes a chip-facing surface facing the first outer surface, the chip-facing surface defines a first opening and a second opening, the first port is used to guide the light emitted by the first optical fiber to pass through the first opening to reach the light-transmitting material, and the second optical fiber is used to receive the scattered light via the second opening.

[0352] Example 24

[0353] For the device according to Embodiment 23, the chip-facing surface is spaced apart from the first outer surface by a gap distance.

[0354] Example 25

[0355] For the device according to any one of Embodiments 1 to 24, it further includes a processor, and the processor is used to determine one or both of the following: determining the particle size of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering assembly or determining the particle size distribution of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering assembly.

[0356] Example 26

[0357] For the device according to Embodiment 25, the processor is used to use autocorrelation to determine one or both of the following: determining the particle size of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering assembly or determining the particle size distribution of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering assembly.

[0358] Example 27

[0359] For the device according to any one of Embodiments 1 to 26, the processing chip is used to form particles encapsulating nucleotides.

[0360] Example 28

[0361] The device according to embodiment 27, wherein the encapsulated nucleotide comprises encapsulated mRNA.

[0362] Example 29

[0363] The device according to any one of embodiments 27 to 28, wherein the nucleotide is encapsulated in a surfactant.

[0364] Example 30

[0365] A device comprising: a processing chip, the processing chip comprising: a first outer surface; a second outer surface; a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber comprising a fluid chamber inlet and a fluid chamber outlet; a light-transmissive material positioned between the first outer surface and the fluid chamber; a plurality of mixing components, each mixing component comprising a mixing stage for mixing a plurality of fluid components to form a fluid mixture, the fluid chamber inlet for receiving the fluid mixture, the fluid mixture comprising particles; a fluid input manifold channel, each of the plurality of mixing components being fluidly coupled to the fluid input manifold channel; a plurality of pressure sensors for sensing the pressure of the fluid components entering the mixing stage of each of the plurality of mixing components; and a dynamic light scattering component, the processing chip being removably positioned relative to the dynamic light scattering component, the dynamic light scattering component for emitting light into the fluid chamber via the light-transmissive material and receiving light scattered from the particles in the fluid mixture in the fluid chamber.

[0366] Example 31

[0367] The device according to embodiment 30, further comprising a processor for receiving data from the dynamic light scattering component.

[0368] Example 32

[0369] The device according to embodiment 31, wherein the processor is configured to determine one or both of the following: determining the particle size of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering component or determining the particle size distribution of the particles in the fluid in the fluid chamber using at least the data from the dynamic light scattering component.

[0370] Example 33

[0371] The device according to any one of embodiments 31 to 32, wherein the processor is further configured to receive data from the plurality of pressure sensors.

[0372] Example 34

[0373] The apparatus according to embodiment 33, wherein the processor is configured to determine whether there is a flow restriction in the mixing level of each of the plurality of mixing components using at least data from a plurality of pressure sensors.

[0374] Example 35

[0375] The apparatus according to any one of embodiments 33 to 34, wherein the processor is configured to further correlate data received from the dynamic light scattering component and the plurality of pressure sensors.

[0376] Example 36

[0377] An apparatus, comprising: a processing chip, the processing chip including: a first outer surface; a second outer surface; a plurality of mixing components, each mixing component including: a first mixing stage having a first mixing outlet, the first mixing stage configured to mix a first plurality of fluid components to form a first fluid mixture and convey the first fluid mixture through the first mixing outlet, the first fluid mixture including particles; and a second mixing stage having a second mixing outlet, the second mixing stage configured to mix a second plurality of fluid components to form a second fluid mixture and convey the second fluid mixture through the second mixing outlet, the second fluid mixture including particles; and a fluid input manifold channel, each of the plurality of mixing components being fluidly coupled to the fluid input manifold channel; a first dynamic light scattering component positioned near the first mixing stage, the first dynamic light scattering component configured to emit light into the first fluid mixture and receive light scattered from particles in the first fluid mixture; and a second dynamic light scattering component positioned near the second mixing stage, the second dynamic light scattering component configured to emit light into the second fluid mixture and receive light scattered from particles in the second fluid mixture; the processing chip being removably positioned relative to the first dynamic light scattering component and the second dynamic light scattering component.

[0378] Example 37

[0379] The apparatus according to embodiment 36, wherein the first dynamic light scattering component is configured to emit light into the first mixing outlet.

[0380] Example 38

[0381] The apparatus according to any one of embodiments 36 to 37, wherein the second dynamic light scattering component is configured to emit light into the second mixing outlet.

[0382] Example 39

[0383] The apparatus according to any one of embodiments 36 to 38, the processing chip further includes a manifold for guiding fluid from the first mixing outlet and the second mixing outlet to a common outlet channel.

[0384] Example 40

[0385] The apparatus according to embodiment 39, the processing chip further includes a fluid chamber positioned between a first outer surface and a second outer surface for receiving a selected one of a first fluid mixture and a second fluid mixture from the common outlet channel; the apparatus further includes: a third dynamic light scattering assembly for emitting light into the fluid chamber and receiving light scattered from particles in the first fluid mixture or the second fluid mixture in the fluid chamber.

[0386] Example 41

[0387] The apparatus according to embodiment 40, further includes a processor for correlating data from the first light scattering assembly and the second light scattering assembly with data from the third dynamic light scattering assembly.

[0388] Example 42

[0389] The apparatus according to any one of embodiments 39 to 41, the processing chip further includes one or more valves for selectively metering the flow rate of fluid from the first mixing outlet and the second mixing outlet to the common outlet channel.

[0390] Example 43

[0391] The apparatus according to any one of embodiments 36 to 42, further includes a plurality of pressure sensors for sensing the pressures of a first fluid component and a second fluid component entering a first mixing stage and a second mixing stage of each of the plurality of mixing components.

[0392] Example 44

[0393] The apparatus according to embodiment 43, further includes a processor for receiving data from the first dynamic light scattering assembly, the second dynamic light scattering assembly, and the plurality of pressure sensors.

[0394] Example 45

[0395] The apparatus according to embodiment 44, the processor for further correlating data from the first dynamic light scattering assembly and the second dynamic light scattering assembly with data from the plurality of pressure sensors.

[0396] Example 46

[0397] A method includes: conveying a fluid through a processing chip to generate encapsulated particles in the fluid; emitting light towards the encapsulated particles via a first optical fiber, the encapsulated particles scattering the emitted light, and the emitted light being transmitted through a light-transmissive material on a first side of the processing chip; receiving the light scattered from the encapsulated particles, the received light being transmitted through the light-transmissive material on the first side of the processing chip, and the received light being received by a second optical fiber that is oriented obliquely with respect to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body located near the processing chip; performing autocorrelation on the received light; and determining any one of the following: determining the particle size of the encapsulated particles using at least autocorrelation, determining the particle size distribution of the encapsulated particles using at least autocorrelation, or determining the particle size and particle size distribution of the encapsulated particles using at least autocorrelation. Conveying the fluid through the processing chip includes conveying the fluid from a fluid input manifold channel of the processing chip to a plurality of mixing components of the processing chip.

[0398] Example 47

[0399] The method according to embodiment 46, wherein the encapsulated particles include encapsulated nucleotides.

[0400] Example 48

[0401] The method according to embodiment 47, wherein the encapsulated nucleotides include encapsulated mRNA.

[0402] Example 49

[0403] The method according to embodiment 48, wherein the encapsulated mRNA includes mRNA encapsulated by at least one delivery carrier molecule.

[0404] Example 50

[0405] The method according to embodiment 49, wherein the at least one delivery carrier molecule includes an amino-lipidated peptoid.

[0406] Example 51

[0407] The method according to any one of embodiments 46 to 50, wherein conveying the fluid through the processing chip to generate encapsulated particles includes conveying two or more fluid components through at least one of the plurality of mixing components to generate the encapsulated particles.

[0408] Example 52

[0409] The method according to any one of embodiments 46 to 51, further comprising monitoring the pressure of the fluid conveyed through the processing chip.

[0410] Example 53

[0411] The method according to embodiment 50 further includes correlating the monitored pressure value with one or both of the determined encapsulated particle size value or the determined encapsulated particle size distribution value.

[0412] Example 54

[0413] The method according to any one of embodiments 52 to 53 further includes: determining that the monitored pressure value falls outside a tolerance range; and stopping the conveyance of fluid through at least a portion of the processing chip in response to determining that the monitored pressure value falls outside the tolerance range.

[0414] Example 55

[0415] The method according to any one of embodiments 46 to 54 further includes: determining that the determined encapsulated particle size or particle size distribution falls outside a tolerance range; and stopping the conveyance of fluid through at least a portion of the processing chip in response to determining that the determined encapsulated particle size or particle size distribution falls outside the tolerance range.

[0416] Example 56

[0417] In the method according to any one of embodiments 46 to 55, the emitted light is emitted along a first axis, the received light is received along a second axis, and the first axis and the second axis intersect at a convergence point, which is positioned within the processing chip.

[0418] Example 57

[0419] In the method according to embodiment 56, the first axis and the second axis together define an angle of inclination.

[0420] Example 58

[0421] In the method according to embodiment 57, the angle of inclination is in the range of about 10 degrees to about 45 degrees.

[0422] Example 59

[0423] A method includes: conveying a fluid through a mixing component of a processing chip to generate a mixture in the fluid that includes encapsulated particles; monitoring a pressure of the fluid conveyed through the mixing component; activating a dynamic light scattering component to determine a particle size or particle size distribution of the encapsulated particles in the fluid, the dynamic light scattering component causing light to scatter off the particles while the fluid is in the processing chip; and correlating the monitored fluid pressure with the determined particle size or particle size distribution. Conveying the fluid through the mixing component includes conveying a first fluid from a fluid input manifold channel to a first mixing component of a plurality of mixing components, the fluid input manifold channel being fluidly coupled to a second mixing component of the plurality of mixing components.

[0424] Example 60

[0425] The method according to embodiment 59, further includes using at least the monitored pressure to regulate the conveyance of the fluid through the mixing component.

[0426] Example 61

[0427] The method according to any one of embodiments 59 to 60, further includes determining that the monitored pressure falls outside a predetermined range and activating the dynamic light scattering component in response to determining that the monitored pressure falls outside the predetermined range.

[0428] Example 62

[0429] The method according to any one of embodiments 59 to 61, activating the dynamic light scattering component includes: emitting light via a first optical fiber toward the encapsulated particles, the encapsulated particles scattering the emitted light, the emitted light being conveyed through a light-transmissive material on a first side of the processing chip, receiving the light scattered from the encapsulated particles, the received light being conveyed through the light-transmissive material on the first side of the processing chip, the received light being received by a second optical fiber that is oriented at an angle relative to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body positioned near the processing chip, performing autocorrelation on the received light, and determining the particle size or particle size distribution of the encapsulated particles using at least the autocorrelation.

[0430] Example 63

[0431] A method, comprising: conveying a fluid from a fluid input manifold channel of a processing chip through a first mixing component of the processing chip to generate a first mixture containing encapsulated particles in the fluid; conveying the fluid from the fluid input manifold channel through a second mixing component of the processing chip to generate a second mixture containing encapsulated particles in the fluid; emitting light towards the encapsulated particles in the first mixture, the particles in the first mixture scattering the emitted light; receiving the light scattered from the encapsulated particles in the first mixture; performing autocorrelation on the received light scattered from the encapsulated particles in the first mixture; using at least the autocorrelation of the received light scattered from the encapsulated particles in the first mixture to determine the particle size or particle size distribution of the encapsulated particles in the first mixture; emitting light towards the encapsulated particles in the second mixture, the particles in the second mixture scattering the emitted light; receiving the light scattered from the encapsulated particles in the second mixture; performing autocorrelation on the received light scattered from the encapsulated particles in the second mixture; and using at least the autocorrelation of the received light scattered from the encapsulated particles in the second mixture to determine the particle size or particle size distribution of the encapsulated particles in the second mixture.

[0432] Example 64

[0433] The method according to embodiment 63, wherein emitting light towards the encapsulated particles in the first mixture, receiving the light scattered from the encapsulated particles in the first mixture, emitting light towards the encapsulated particles in the second mixture, and receiving the light scattered from the encapsulated particles in the second mixture are performed by a single dynamic light scattering component.

[0434] Example 65

[0435] The method according to embodiment 64, further comprising: selectively conveying the first mixture to a fluid chamber of a single dynamic light scattering component, and emitting light towards the encapsulated particles in the first mixture and receiving the light scattered from the encapsulated particles in the first mixture are performed when the first mixture is in the fluid chamber; and selectively conveying the second mixture to the fluid chamber, and emitting light towards the encapsulated particles in the second mixture and receiving the light scattered from the encapsulated particles in the second mixture are performed when the second mixture is in the fluid chamber.

[0436] Example 66

[0437] The method according to any one of embodiments 63 to 64 further comprises: activating a first dynamic light scattering component to emit light towards the encapsulated particles in the first mixture and receive the light scattered from the encapsulated particles in the first mixture; and activating a second dynamic light scattering component, separate from the first dynamic light scattering component, to emit light towards the encapsulated particles in the second mixture and receive the light scattered from the encapsulated particles in the second mixture.

[0438] Example 67

[0439] The method according to embodiment 66 further comprises: selectively delivering the first mixture to a fluid chamber of a third dynamic light scattering component; emitting light towards the encapsulated particles in the first mixture while the first mixture is in the fluid chamber; receiving the light scattered from the encapsulated particles in the first mixture while the first mixture is in the fluid chamber; selectively delivering the second mixture to the fluid chamber; emitting light towards the encapsulated particles in the second mixture while the second mixture is in the fluid chamber; and receiving the light scattered from the encapsulated particles in the second mixture while the second mixture is in the fluid chamber.

[0440] Example 68

[0441] The method according to embodiment 67 further comprises passing the first mixture and the second mixture through a manifold, the first mixture and the second mixture passing through the manifold before reaching the fluid chamber.

[0442] Example 69

[0443] An apparatus includes: a body that includes a first port and a second port and that is capable of being positioned adjacent to a first outer surface of a processing chip; a first optical fiber coupled to the first port of the body, the first optical fiber for emitting light, the first port for guiding light emitted by the first optical fiber through a light-transmissive material of the processing chip and into a fluid chamber of the processing chip; a focusing lens supported by the body, the focusing lens positioned and configured to focus light emitted by the first optical fiber; a second optical fiber coupled to the second port of the body, the second optical fiber at the second port being oriented obliquely with respect to the first optical fiber at the first port, the second optical fiber for receiving light scattered by particles in a fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber; a filter supported by the body, the filter positioned and configured to filter light scattered by particles in a fluid in the fluid chamber; and a processing chip removably positioned relative to the body, the processing chip including: a first outer surface, a second outer surface, a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet, a light-transmissive material positioned between the first outer surface and the fluid chamber, a plurality of mixing components, and a fluid input manifold channel fluidly coupled to each of the plurality of mixing components.

[0444] Example 70

[0445] The apparatus according to embodiment 69, further comprising a base having a processing chip mount for removably receiving the processing chip, the body being positioned adjacent to the processing chip mount.

[0446] Example 71

[0447] The apparatus according to any one of embodiments 69 to 70, the body for orienting the first optical fiber along an axis perpendicular to an outer surface of the processing chip.

[0448] Example 72

[0449] The apparatus according to any one of embodiments 69 to 71, the body for orienting the second optical fiber along an axis that is oriented obliquely with respect to an outer surface of the processing chip.

[0450] Example 73

[0451] The apparatus according to any one of embodiments 69 to 72, further comprising a processor configured to receive data indicative of light received by the second optical fiber and to selectively direct fluid from the fluid input manifold channel to one of the plurality of mixing components based on the data.

[0452] Example 74

[0453] An apparatus, comprising: a processing chip mount for removably receiving a processing chip; a body including a first port and a second port, the body being fixedly secured relative to the processing chip mount, the processing chip mount for removably receiving the processing chip between the body and the processing chip mount; a first optical fiber coupled to the first port of the body, the first optical fiber for emitting light, the first port for guiding the light emitted by the first optical fiber through a light-transmissive material of the processing chip received by the processing chip mount and into a fluid chamber of the processing chip; a second optical fiber coupled to the second port of the body, the second optical fiber at the second port being inclinedly oriented relative to the first optical fiber at the first port, the second optical fiber for receiving light scattered by particles in a fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber; and a processing chip removably received in the processing chip mount, the processing chip including: a first outer surface; a second outer surface; a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet; a light-transmissive material and a plurality of mixing components positioned between the first outer surface and the fluid chamber, and a fluid input manifold channel fluidly coupled to each of the plurality of mixing components.

[0454] Example 75

[0455] The apparatus according to embodiment 74, wherein the processing chip is configured to form a therapeutic composition.

[0456] Example 76

[0457] The apparatus according to embodiment 75, wherein the therapeutic composition comprises a fluid containing particles.

[0458] Example 77

[0459] The apparatus according to embodiment 76, wherein the first port is for guiding the light emitted by the first optical fiber through the light-transmissive material of the processing chip to the fluid of the therapeutic composition.

[0460] Example 78

[0461] The apparatus according to embodiment 77, wherein the second optical fiber is for receiving the light scattered by the particles of the therapeutic composition.

[0462] Example 79

[0463] The device according to embodiment 78 further includes a processor configured to determine one or both of the following: the particle size of particles in the therapeutic composition using light scattered at least by the particles of the therapeutic composition, or the particle size distribution of particles in the therapeutic composition using light scattered at least by the particles of the therapeutic composition.

[0464] Example 80

[0465] In the device according to embodiment 79, the processor is configured to selectively direct fluid from a fluid input manifold channel to one of a plurality of mixing components based on one or both of the determined particle size of particles in the therapeutic composition or the particle size distribution of particles in the therapeutic composition.

[0466] Example 81

[0467] A device includes: a processing chip including: a fluid chamber having a fluid chamber inlet and a fluid chamber outlet, and a light-transmissive material adjacent to the fluid chamber; a dynamic light scattering assembly, the processing chip being removably positioned relative to the dynamic light scattering assembly, the dynamic light scattering assembly configured to direct light through the light-transmissive material and into the fluid chamber, the dynamic light scattering assembly further configured to receive light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber, and thereby capture light scattering data; and a processor configured to determine the viscosity of the fluid in the fluid chamber based on the captured light scattering data, the processor further configured to determine one or both of the particle size or particle size distribution of particles in the fluid based on the captured light scattering data, the processing chip further including a plurality of mixing components, each of the plurality of mixing components having a plurality of inlets and outlets, each of the plurality of mixing components configured to form a fluid mixture from the plurality of inlets and convey the mixture through the outlet, the processing chip further including a fluid input port and a fluid input manifold channel, the plurality of inlets of the plurality of mixing components being fluidly coupled to the fluid input manifold channel.

[0468] Example 82

[0469] In the device according to embodiment 81, the processing chip further includes: a first channel, the fluid chamber inlet being configured to receive a first fluid from the first channel, and a second channel, the fluid chamber inlet being further configured to receive a second fluid from the second channel.

[0470] Example 83

[0471] In the device according to embodiment 82, the first fluid includes a therapeutic composition.

[0472] Example 84

[0473] The device according to Example 83, wherein the therapeutic composition comprises at least some of the particles in the granules.

[0474] Example 85

[0475] The device according to Example 84, wherein the particles of the therapeutic composition comprise mRNA.

[0476] Example 86

[0477] The device according to any one of Examples 82 to 85, wherein the second fluid comprises at least some of the particles in the granules.

[0478] Example 87

[0479] The device according to Example 86, wherein the particles of the second fluid comprise beads.

[0480] Example 88

[0481] The device according to Example 87, wherein the first fluid comprises a therapeutic composition, and the therapeutic composition comprises particles.

[0482] Example 89

[0483] The device according to Example 88, wherein the particles of the therapeutic composition have a first diameter, and the beads have a second diameter different from the first diameter.

[0484] Example 90

[0485] The device according to Example 89, wherein the second diameter is greater than the first diameter.

[0486] Example 91

[0487] The device according to any one of Examples 82 to 90, wherein the first fluid contains particles of a first type, and the second fluid contains particles of a second type.

[0488] Example 92

[0489] The device according to Example 91, wherein the processor is configured to determine one or both of the particle size or particle size distribution of the particles of the first type in the first fluid based on the light scattered by the particles of the first type and the particles of the second type.

[0490] Example 93

[0491] The device according to Embodiment 92, wherein the second type of particles has a known particle size.

[0492] Example 94

[0493] The device according to any one of Embodiments 82 to 93, wherein the second fluid comprises a diluent.

[0494] Example 95

[0495] The device according to Embodiment 94, wherein the processing chip is configured to selectively add discrete amounts of diluent to the first fluid in a sequence.

[0496] Example 96

[0497] The device according to Embodiment 95, wherein the processing chip includes at least one valve to selectively control the delivery of the diluent to the first fluid.

[0498] Example 97

[0499] The device according to any one of Embodiments 95 to 96, wherein the processing chip further includes at least one pump to selectively drive the movement of the diluent.

[0500] Example 98

[0501] The device according to any one of Embodiments 94 to 97, wherein the processor is configured to: during a sequence of adding discrete amounts of diluent to the first fluid, track the autocorrelation of the captured light scattering data and determine one or both of the particle size or the particle size distribution of the particles in the fluid based on the tracked autocorrelation.

[0502] Example 99

[0503] The device according to any one of Embodiments 94 to 98, wherein the processing chip further includes a mixing chamber to mix the diluent with the first fluid.

[0504] Example 100

[0505] The device according to Embodiment 99, wherein the mixing chamber is positioned adjacent to the fluid chamber.

[0506] Example 101

[0507] The device according to any one of Embodiments 99 to 100, wherein the processing chip further includes a first pump and a second pump, the first pump and the second pump being configured to be alternately activated to drive the combination of the diluent and the first fluid back and forth through the mixing chamber.

[0508] Example 102

[0509] A method includes: conveying a fluid mixture through a processing chip, the fluid mixture containing particles; emitting light towards the fluid mixture via a first optical fiber, the particles in the fluid mixture scattering the emitted light; receiving the light scattered from the particles in the fluid mixture, the received light being received by a second optical fiber that is oriented obliquely with respect to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body located near the processing chip; performing autocorrelation on the received light; using at least the autocorrelation to determine the viscosity of the fluid mixture; and determining any one of the following: using at least the autocorrelation to determine the particle size of the particles in the fluid mixture, using at least the autocorrelation to determine the particle size distribution of the particles in the fluid mixture, or using at least the autocorrelation to determine the particle size and particle size distribution of the particles in the fluid mixture: Conveying the fluid through the processing chip includes: conveying a first fluid component from a fluid input manifold channel to a first mixing assembly, the fluid input manifold channel being fluidly coupled to a second mixing assembly, the first fluid component including at least some of the particles, conveying a second fluid component to the first mixing assembly, and mixing the first fluid component and the second fluid component together to form a fluid mixture.

[0510] Example 103

[0511] The method according to embodiment 102, wherein the particles of the first fluid component include therapeutic particles.

[0512] Example 104

[0513] The method according to embodiment 103, wherein the therapeutic particles include mRNA.

[0514] Example 105

[0515] The method according to embodiment 104, wherein the mRNA is encapsulated in a delivery vehicle.

[0516] Example 106

[0517] The method according to any one of embodiments 102 to 105, wherein the second fluid component includes at least some of the particles.

[0518] Example 107

[0519] The method according to embodiment 106, wherein the particles of the second fluid component include beads.

[0520] Example 108

[0521] According to the method described in any one of embodiments 106 to 107, the particles of the first fluid component have a first diameter, and the particles of the second fluid component have a second diameter different from the first diameter.

[0522] Example 109

[0523] According to the method described in any one of embodiments 108, the second diameter is greater than the first diameter.

[0524] Example 110

[0525] According to the method described in any one of embodiments 106 to 109, the particles of the first fluid component include a first type of particles, and the particles of the second fluid component include a second type of particles different from the first type of particles.

[0526] Example 111

[0527] According to the method described in any one of embodiments 106 to 110, receiving light scattered from the particles in the fluid mixture includes: receiving light scattered by the particles of the first fluid component and receiving light scattered by the particles of the second fluid component.

[0528] Example 112

[0529] According to the method described in embodiment 111, determining any of the following: determining the particle size of the particles in the fluid mixture using at least autocorrelation, determining the particle size distribution of the particles in the fluid mixture using at least autocorrelation, or determining the particle size and particle size distribution of the particles in the fluid mixture using at least autocorrelation, includes determining any of the following: determining the particle size of the particles of the first fluid component using at least autocorrelation, determining the particle size distribution of the particles of the first fluid component using at least autocorrelation, or determining the particle size and particle size distribution of the particles of the first fluid component using at least autocorrelation.

[0530] Example 113

[0531] According to the method described in any one of embodiments 102 to 112, the second fluid component contains a diluent.

[0532] Example 114

[0533] According to the method described in embodiment 113, conveying the fluid mixture through the processing chip further includes adding discrete amounts of diluent to the first fluid component in sequence.

[0534] Example 115

[0535] The method according to Embodiment 114 further includes repeating emitting light, receiving light, and performing autocorrelation each time a discrete amount of diluent is added to the first fluid component in sequence.

[0536] Example 116

[0537] The method according to Embodiment 115 further includes tracking the autocorrelation throughout each repetition of emitting light, receiving light, and performing autocorrelation each time a discrete amount of diluent is added to the first fluid component in sequence.

[0538] Example 117

[0539] The method according to Embodiment 116 of using at least autocorrelation to determine the viscosity of a fluid mixture includes using the tracked autocorrelation to determine the viscosity of the fluid mixture.

[0540] Example 118

[0541] The method according to any one of Embodiments 116 to 117 of determining any of the following: using at least autocorrelation to determine the particle size of particles in a fluid mixture, using at least autocorrelation to determine the particle size distribution of particles in a fluid mixture, or using at least autocorrelation to determine the particle size and particle size distribution of particles in a fluid mixture includes determining any of the following: using the tracked autocorrelation to determine the particle size of particles in a fluid mixture, using the tracked autocorrelation to determine the particle size distribution of particles in a fluid mixture, or using the tracked autocorrelation to determine the particle size and particle size distribution of particles in a fluid mixture.

[0542] Example 119

[0543] The discrete amount of diluent added to the first fluid component is the same amount of diluent each time a discrete amount of diluent is added to the first fluid component in sequence according to the method of any one of Embodiments 114 to 118.

[0544] Example 120

[0545] Mixing the first fluid component and the second fluid component together to form a fluid mixture according to the method of any one of Embodiments 102 to 119 includes alternately activating at least two pumps to drive the first fluid component and the second fluid component back and forth through the mixing chamber of the processing chip.

[0546] Example 121

[0547] An apparatus includes: a processing chip, the processing chip including: a first fluid input port, a first fluid input manifold fluidly coupled to the first fluid input port; a second fluid input port, a second fluid input manifold fluidly coupled to the second fluid input port; a plurality of mixing components, each mixing component including: a first valve fluidly coupled to the first input manifold, a first inlet fluidly coupled to the first valve, a second valve fluidly coupled to the second input manifold, a second inlet fluidly coupled to the second valve, an outlet; the mixing components being configured to form a fluid mixture from the first inlet and the second inlet and convey the mixture out through the outlet; one or more measurement features that are operable to detect one or more characteristics of the mixture; and a processor configured to activate the first valve and the second valve of the plurality of mixing components based on data from the one or more measurement features.

[0548] VII. Miscellaneous

[0549] The foregoing description is provided to enable those skilled in the art to practice the various configurations described herein. While the subject technology has been described specifically with reference to various drawings and configurations, it should be understood that these are for illustrative purposes only and should not be considered as limiting the scope of the subject technology.

[0550] There may be many other ways to implement the subject technology. Without departing from the scope of the subject technology, the various functions and elements described herein may be different from those shown. Various modifications to these specific implementations will be apparent to those skilled in the art, and the general principles defined herein may be applied to other specific implementations. Thus, many changes and modifications may be made to the subject technology by those of ordinary skill in the art without departing from the scope of the subject technology. For example, a different number of a given module or unit may be employed, a different type of a given module or unit may be employed, a given module or unit may be added, or a given module or unit may be omitted.

[0551] When a feature or element is referred to herein as being "on" another feature or element, it can be directly on the other feature or element or intervening features and / or elements may be present. In contrast, when a feature or element is referred to as being "directly on" another feature or element, no intervening features or elements are present. When a feature or element is referred to as being "connected", "attached" or "coupled" to another feature or element, it can be directly connected, attached or coupled to the other feature or element, or intervening features or elements may be present. In contrast, when a feature or element is referred to as being "directly connected", "directly attached" or "directly coupled" to another feature or element, no intervening features or elements are present. Although described or shown with respect to one embodiment, the features and elements so described or shown can be applied to other embodiments. Those skilled in the art will also understand that references to a structure or feature that is "adjacent" to another feature can have portions that overlap or are beneath the adjacent feature.

[0552] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. For example, as used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items and may be abbreviated as " / ".

[0553] Spatial relative terms, such as "below", "beneath", "lower", "above", "upper", etc., may be used herein for ease of description to describe the relationship of one element or feature to another element or feature as illustrated. It should be understood that the spatial relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if the device in the figures is inverted, an element described as "below" or "beneath" another element or feature will then be oriented "above" the other element or feature. Thus, the term "below" can encompass both an orientation of above and below. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatial relative descriptors used herein are to be interpreted accordingly. Similarly, the terms "upward", "downward", "vertical", "horizontal", etc. are used herein for explanatory purposes only unless specifically indicated otherwise.

[0554] As used herein, the term "perpendicular" should be understood to include an arrangement in which two objects, axes, planes, surfaces, or other objects are oriented such that the two objects, axes, planes, surfaces, or other objects together define a 90-degree angle. The term "perpendicular" as used herein should also be understood to include an arrangement in which two objects, axes, planes, surfaces, or other objects are oriented such that the two objects, axes, planes, surfaces, or other objects together define an approximate 90-degree angle (e.g., an angle in the range from 85 degrees to 90 degrees). Thus, the term "perpendicular" as used herein should not be understood to necessarily require an arrangement in which two objects, axes, planes, surfaces, or other objects are oriented such that the two objects, axes, planes, surfaces, or other objects together define an exact 90-degree angle.

[0555] Although the terms "first" and "second" may be used herein to describe various features / elements (including steps), these features / elements should not be limited by these terms, unless the context indicates otherwise. These terms may be used to distinguish one feature / element from another. Thus, without departing from the teachings of the present invention, the first feature / element discussed below may be referred to as the second feature / element, and similarly, the second feature / element discussed below may be referred to as the first feature / element.

[0556] Throughout the specification and the following claims, unless the context requires otherwise, the words "comprise", "comprising", "include" mean that various components may be used together in a method and an article (e.g., a composition and an apparatus including a device and a method). For example, the term "comprising" will be understood to imply the inclusion of any stated element or step, but not the exclusion of any other element or step. Generally, any of the apparatus and methods described herein should be understood to be inclusive, but the whole or a subset of components and / or steps may alternatively be exclusive and may be expressed as "consisting of various components, steps, sub-components, or sub-steps" or alternatively "consisting essentially of various components, steps, sub-components, or sub-steps".

[0557] As used herein in the specification and claims, including as used in the examples, and unless otherwise expressly specified, all numbers may be read as if prefaced by the word "about" or "approximately", even if the term does not expressly appear. When describing magnitudes and / or positions, the phrase "about" or "approximately" may be used to indicate that the described value and / or position is within a reasonable expectation of the value and / or position. For example, a numerical value may have a value of + / −0.1%, + / −1%, + / −2%, + / −5%, + / −10%, etc. of the stated value (or range of values). Any numerical value given herein should also be understood to include about or approximately that value, unless the context otherwise indicates. For example, if the value "10" is disclosed, then "about 10" is also disclosed. Any numerical range recited herein is intended to include all sub-ranges subsumed therein.

[0558] It should also be understood that when a value is disclosed, then "less than or equal to" that value, "greater than or equal to" that value, and the possible ranges between values are also disclosed, as would be appropriately understood by one of ordinary skill in the art. For example, if the value "X" is disclosed, then "less than or equal to X" and "greater than or equal to X" are also disclosed (e.g., where X is a numerical value). It should also be understood that throughout the application, data is provided in a variety of different formats, and that this data represents ranges of endpoints and starting points and any combination of data points. For example, if a particular data point "10" and a particular data point "15" are disclosed, then it should be understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 and between 10 and 15 are all considered to be disclosed. It should also be understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0559] As used herein, the terms "system", "apparatus", and "device" may be understood to be interchangeable. A system, apparatus, and device may each include a plurality of components having various structural and / or functional relationships to one another.

[0560] Some forms of the examples described herein may be implemented using a computer system, where the computer system may include at least one processor communicating with a number of peripheral devices via a bus subsystem. Forms of the examples described herein implemented using a computer system may be implemented using a general purpose computer programmed to perform the methods described herein. Alternatively, forms of the examples described herein implemented using a computer system may be implemented using a special purpose computer constructed of hardware arranged to perform the methods described herein. Forms of the examples described herein may also be implemented using a combination of at least one general purpose computer and at least one special purpose computer.

[0561] In an implementation using a computer system, each processor may include a central processing unit (CPU) of the computer system, a microprocessor, an application specific integrated circuit (ASIC), other types of hardware components, and combinations thereof. The computer system may include more than one type of processor. The peripheral devices of the computer system may include a storage subsystem (including, for example, memory devices and file storage subsystems), user interface input devices, user interface output devices, and network interface subsystems. The input and output devices may allow a user to interact with the computer system. The network interface subsystem may provide an interface to an external network, including an interface to a corresponding interface device in other computer systems. The user interface input devices may include a keyboard; pointing devices such as a mouse, trackball, touchpad, or graphics tablet; a scanner; a touchscreen incorporated into a display; audio input devices such as a voice recognition system and a microphone; and other types of input devices. Generally, the use of the term "input device" is intended to include all possible types of devices and ways of inputting information into the computer system.

[0562] In an implementation using a computer system, the user interface output devices may include a display subsystem, a printer, a fax machine, or a non-visual display such as an audio output device. The display subsystem may include a cathode ray tube (CRT), a flat panel device such as a liquid crystal display (LCD), a projection device, or some other mechanism for creating visual images. The display subsystem may also provide a non-visual display, such as an audio output device. Generally, the use of the term "output device" is intended to include all possible types of devices and ways of outputting information from the computer system to a user or to another machine or computer system.

[0563] In an implementation using a computer system, the storage subsystem may store programming and data constructs that provide some or all of the functionality of the modules and methods described herein. These software modules may typically be executed by the processor of the computer system alone or in combination with other processors. The memory used in the storage subsystem may include multiple memories, including a main random access memory (RAM) for storing instructions and data during program execution and a read only memory (ROM) in which fixed instructions are stored. The file storage subsystem may provide persistent storage of program and data files, and may include a hard disk drive, a floppy disk drive together with an associated removable medium, a CD-ROM drive, an optical disk drive, or a removable media cartridge. The modules implementing the functions of certain specific implementations may be stored in the storage subsystem by the file storage subsystem, or stored in other machines accessible by the processor.

[0564] In an implementation using a computer system, the computer system itself can be of various types, including personal computers, portable computers, workstations, computer terminals, network computers, televisions, mainframes, server farms, a widely distributed group of loosely networked computers, or any other data processing system or user device. Due to the ever-changing nature of computers and networks, the examples of computer systems described herein are only intended to be specific examples for illustrating the disclosed technology. Many other configurations of computer systems with more or fewer components than those described herein are possible.

[0565] As an article of manufacture rather than a method, a non-transitory computer-readable medium (CRM) can be loaded with program instructions executable by a processor. The program instructions, when executed, implement one or more of the above-described computer-implemented methods. Alternatively, the program instructions can be loaded onto a non-transitory CRM and, when combined with appropriate hardware, become components of one or more of the computer-implemented systems practicing the disclosed methods.

[0566] The underlined and / or italicized headings and subheadings are used for convenience only, do not limit the claimed subject matter, and are not to be construed as part of the description of the claimed subject matter. All structural and functional equivalents of the elements of the various specific implementations described throughout this disclosure that are known or later become known to those of ordinary skill in the art are hereby expressly incorporated by reference and are intended to be covered by the claimed subject matter. Moreover, any disclosure herein is not intended to be dedicated to the public regardless of whether such disclosure is expressly recited above.

[0567] It should be understood that all combinations of the foregoing concepts and additional concepts discussed in greater detail below (assuming such concepts are not mutually inconsistent) are considered to be part of the inventive subject matter disclosed herein. Specifically, all combinations of the claimed subject matter appearing at the end of this disclosure are considered to be part of the inventive subject matter disclosed herein.

Claims

1. An apparatus, comprising: a processing chip, the processing chip comprising: a first outer surface, a second outer surface, a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet, a light-transmissive material positioned between the first outer surface and the fluid chamber, a plurality of mixing components, each of the plurality of mixing components having a plurality of inlets and an outlet, each of the plurality of mixing components configured to form a mixture of fluids from the plurality of inlets and convey the mixture through the outlet, and a fluid input manifold channel, at least one inlet of each of the plurality of mixing components being fluidly coupled to the fluid input manifold channel; and a dynamic light scattering component, the processing chip being removably positioned relative to the dynamic light scattering component, the dynamic light scattering component comprising: a body including a first port and a second port, the body being positioned adjacent to the first outer surface, a first optical fiber coupled to the first port of the body, the first optical fiber for emitting light, the first port for guiding the light emitted by the first optical fiber through the light-transmissive material and into the fluid chamber, and a second optical fiber coupled to the second port of the body, the second optical fiber at the second port being oriented obliquely relative to the first optical fiber at the first port, the second optical fiber for receiving light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber.

2. The apparatus according to claim 1, wherein the fluid chamber has a cylindrical shape including a circular upper inner surface, a circular lower inner surface, and an inner sidewall extending from the circular upper inner surface to the circular lower inner surface.

3. The apparatus according to claim 2, wherein the fluid chamber inlet is positioned in a region of the inner sidewall adjacent to the circular lower inner surface.

4. The apparatus according to any one of claims 2 to 3, wherein the fluid chamber outlet is positioned in a region of the inner sidewall adjacent to the circular upper inner surface.

5. The apparatus according to any one of claims 1 to 4, wherein at least one of the plurality of mixing components includes a first mixing stage for mixing a first plurality of fluid components to form a first fluid mixture, the fluid chamber inlet for receiving the first fluid mixture.

6. The apparatus according to claim 5, wherein the first mixing stage includes a first mixing inlet, a second mixing inlet, and a first mixing outlet, the first mixing inlet for receiving a first fluid component, the second mixing inlet for receiving a second fluid component, the first mixing outlet for outputting the first fluid mixture, the first fluid mixture comprising at least the first fluid component and the second fluid component.

7. The apparatus according to claim 6, wherein the processing chip further comprises: A first pressure sensor configured to sense a pressure of the first fluid component entering the first mixing inlet, and A second pressure sensor configured to sense a pressure of the second fluid component entering the second mixing inlet.

8. The apparatus according to claim 7, further comprising a processor configured to receive data from the dynamic light scattering assembly, the first pressure sensor, and the second pressure sensor.

9. The apparatus according to claim 8, wherein the processor is configured to further correlate the data received from the dynamic light scattering assembly, the first pressure sensor, and the second pressure sensor.

10. The apparatus according to any one of claims 6 to 9, wherein the processing chip further comprises an additional fluid passage fluidly coupled to the first mixing outlet.

11. The apparatus according to claim 10, wherein the processing chip is configured to provide conveyance of fluid from the first mixing outlet to the additional fluid passage, the fluid chamber inlet, or a combination of the additional fluid passage and the fluid chamber inlet.

12. The apparatus according to any one of claims 10 to 11, wherein the processing chip is configured to provide conveyance of fluid from the additional fluid passage to the fluid chamber inlet via the first mixing outlet.

13. The apparatus according to any one of claims 5 to 12, wherein at least one of the plurality of mixing assemblies further comprises a second mixing stage having a second mixing outlet configured to mix a second plurality of fluid components to form a second fluid mixture, and the fluid chamber inlet is configured to receive the second fluid mixture from the second mixing outlet.

14. The apparatus according to claim 13, wherein the processing chip further comprises at least one valve configured to regulate a flow rate of fluid from the first mixing outlet and the second mixing outlet to the fluid chamber inlet such that the fluid chamber inlet selectively receives only one of the first fluid mixture or the second fluid mixture at a time.

15. The apparatus according to any one of claims 13 to 14, wherein the processing chip further comprises a manifold configured to direct fluid from the first mixing outlet and the second mixing outlet to the fluid chamber inlet.

16. The apparatus according to any one of claims 1 to 15, wherein the processing chip has a square shape including four corners, and the dynamic light scattering assembly is positioned at one of the four corners.

17. The apparatus according to any one of claims 1 to 16, wherein the dynamic light scattering assembly further comprises a collimator located in the first port, the collimator being interposed between an end of the first optical fiber and the first outer surface.

18. The apparatus according to claim 17, wherein the first port further comprises a focusing volume interposed between the collimator and the first outer surface.

19. The apparatus according to claim 18, wherein the focusing volume defines a conical shape.

20. The device according to any one of claims 18 to 19, wherein the dynamic light scattering assembly further comprises a focusing lens interposed between the collimator and the focusing volume.

21. The device according to any one of claims 1 to 20, wherein the dynamic light scattering assembly further comprises a filter located in the second port, the filter being interposed between the end of the second optical fiber and the first outer surface.

22. The device according to claim 21, wherein the body further defines a channel interposed between the filter and the first outer surface.

23. The device according to any one of claims 1 to 22, wherein the body includes a chip-facing surface facing the first outer surface, the chip-facing surface defining a first opening and a second opening, the first port being for guiding light emitted by the first optical fiber through the first opening to the light-transmissive material, and the second optical fiber being for receiving scattered light via the second opening.

24. The device according to claim 23, wherein the chip-facing surface is spaced apart from the first outer surface by a gap distance.

25. The device according to any one of claims 1 to 24, further comprising a processor configured to determine one or both of the following: determining the particle size of particles in the fluid in the fluid chamber using at least data from the dynamic light scattering assembly or determining the particle size distribution of particles in the fluid in the fluid chamber using at least data from the dynamic light scattering assembly.

26. The device according to claim 25, wherein the processor is configured to use autocorrelation to determine one or both of the following: determining the particle size of particles in the fluid in the fluid chamber using at least data from the dynamic light scattering assembly or determining the particle size distribution of particles in the fluid in the fluid chamber using at least data from the dynamic light scattering assembly.

27. The device according to any one of claims 1 to 26, wherein the processing chip is configured to form particles encapsulating nucleotides.

28. The device according to claim 27, wherein the encapsulated nucleotides include encapsulated mRNA.

29. The device according to any one of claims 27 to 28, wherein the nucleotides are encapsulated in a surfactant.

30. A device comprising: a processing chip, the processing chip comprising: a first outer surface, a second outer surface, a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet, a light-transmissive material positioned between the first outer surface and the fluid chamber, a plurality of mixing assemblies, each mixing assembly including a mixing stage for mixing a plurality of fluid components to form a fluid mixture, the fluid chamber inlet being for receiving the fluid mixture, the fluid mixture containing particles, a fluid input manifold channel, each of the plurality of mixing assemblies being fluidly coupled to the fluid input manifold channel A plurality of pressure sensors configured to sense the pressure of a fluid component at a mixing stage of each of the plurality of mixing assemblies as the fluid component enters the mixing assemblies; and A dynamic light scattering assembly, with the processing chip removably positioned relative to the dynamic light scattering assembly, the dynamic light scattering assembly configured to emit light into the fluid chamber through the transmissive material and receive light scattered by particles in the fluid mixture within the fluid chamber.

31. The apparatus of claim 30, further comprising a processor configured to receive data from the dynamic light scattering assembly.

32. The apparatus of claim 31, wherein the processor is configured to determine one or both of: determining a particle size of particles in the fluid within the fluid chamber using at least data from the dynamic light scattering assembly or determining a particle size distribution of particles in the fluid within the fluid chamber using at least data from the dynamic light scattering assembly.

33. The apparatus of any one of claims 31 to 32, wherein the processor is further configured to receive data from the plurality of pressure sensors.

34. The apparatus of claim 33, wherein the processor is configured to use at least data from the plurality of pressure sensors to determine whether there is a flow restriction at the mixing stage of each of the plurality of mixing assemblies.

35. The apparatus of any one of claims 33 to 34, wherein the processor is configured to further correlate the data received from the dynamic light scattering assembly and the plurality of pressure sensors.

36. An apparatus comprising: A processing chip, the processing chip comprising: A first outer surface, A second outer surface, A plurality of mixing assemblies, each mixing assembly comprising: A first mixing stage having a first mixing outlet, the first mixing stage configured to mix a first plurality of fluid components to form a first fluid mixture and convey the first fluid mixture through the first mixing outlet, the first fluid mixture containing particles, and A second mixing stage having a second mixing outlet, the second mixing stage configured to mix a second plurality of fluid components to form a second fluid mixture and convey the second fluid mixture through the second mixing outlet, the second fluid mixture containing particles, and A fluid input manifold channel, with each of the plurality of mixing assemblies fluidly coupled to the fluid input manifold channel; A first dynamic light scattering assembly positioned near the first mixing stage, the first dynamic light scattering assembly configured to emit light into the first fluid mixture and receive light scattered by particles in the first fluid mixture; and A second dynamic light scattering assembly positioned near the second mixing stage, the second dynamic light scattering assembly configured to emit light into the second fluid mixture and receive light scattered by particles in the second fluid mixture; The processing chip is removably positioned relative to the first dynamic light scattering assembly and the second dynamic light scattering assembly.

37. The apparatus according to claim 36, wherein the first dynamic light scattering component is configured to emit light into the first mixing outlet.

38. The apparatus according to any one of claims 36 to 37, wherein the second dynamic light scattering component is configured to emit light into the second mixing outlet.

39. The apparatus according to any one of claims 36 to 38, wherein the processing chip further includes a manifold configured to direct fluid from the first mixing outlet and the second mixing outlet to a common outlet channel.

40. The apparatus according to claim 39, wherein the processing chip further includes a fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber configured to receive a selected one of the first fluid mixture and the second fluid mixture from the common outlet channel; The apparatus further comprises: a third dynamic light scattering component configured to emit light into the fluid chamber and receive light scattered by particles in the first fluid mixture or the second fluid mixture in the fluid chamber.

41. The apparatus according to claim 40, further comprising a processor configured to correlate data from the first light scattering component and the second light scattering component with data from the third dynamic light scattering component.

42. The apparatus according to any one of claims 39 to 41, wherein the processing chip further includes one or more valves to selectively meter the flow of fluid from the first mixing outlet and the second mixing outlet to the common outlet channel.

43. The apparatus according to any one of claims 36 to 42, further comprising a plurality of pressure sensors configured to sense the pressures of the first fluid component and the second fluid component entering the first mixing stage and the second mixing stage of each of the plurality of mixing components.

44. The apparatus according to claim 43, further comprising a processor configured to receive data from the first dynamic light scattering component, the second dynamic light scattering component, and the plurality of pressure sensors.

45. The apparatus according to claim 44, wherein the processor is configured to further correlate the data from the first dynamic light scattering component and the second dynamic light scattering component with the data from the plurality of pressure sensors.

46. A method, the method comprises: transferring a fluid through a processing chip to generate encapsulated particles in the fluid; emitting light via a first optical fiber toward the encapsulated particles, the encapsulated particles scattering the emitted light, the emitted light being transmitted through a light-transmissive material on a first side of the processing chip; receiving the light scattered from the encapsulated particles, the received light being transmitted through the light-transmissive material on the first side of the processing chip, the received light being received by a second optical fiber oriented obliquely with respect to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body located near the processing chip; performing autocorrelation on the received light; and Determine any of the following: determining the particle size of the encapsulated particles using at least the autocorrelation, determining the particle size distribution of the encapsulated particles using at least the autocorrelation, or determining the particle size and particle size distribution of the encapsulated particles using at least the autocorrelation, Conveying fluid through the processing chip includes conveying the fluid from a fluid input manifold channel of the processing chip to a plurality of mixing components of the processing chip.

47. The method according to claim 46, wherein the encapsulated particles comprise encapsulated nucleotides.

48. The method according to claim 47, wherein the encapsulated nucleotides comprise encapsulated mRNA.

49. The method according to claim 48, wherein the encapsulated mRNA comprises mRNA encapsulated by at least one delivery carrier molecule.

50. The method according to claim 49, wherein the at least one delivery carrier molecule comprises an amino-lipidated peptidomimetic.

51. The method according to any one of claims 46 to 50, wherein conveying fluid through the processing chip to generate encapsulated particles in the fluid includes conveying two or more fluid components through at least one of the plurality of mixing components to generate the encapsulated particles.

52. The method according to any one of claims 46 to 51, further comprising monitoring the pressure of the fluid conveyed through the processing chip.

53. The method according to claim 50, further comprising correlating the monitored pressure value with one or both of the determined encapsulated particle size value or the determined encapsulated particle size distribution value.

54. The method according to any one of claims 52 to 53, further comprising: determining that the monitored pressure value falls outside a tolerance range; and responsive to determining that the monitored pressure value falls outside a tolerance range, stopping conveying fluid through at least a portion of the processing chip.

55. The method according to any one of claims 46 to 54, further comprising: determining that the determined encapsulated particle size or particle size distribution falls outside a tolerance range; and responsive to determining that the determined encapsulated particle size or particle size distribution falls outside a tolerance range, stopping conveying fluid through at least a portion of the processing chip.

56. The method according to any one of claims 46 to 55, wherein the emitted light is emitted along a first axis and the received light is received along a second axis, and the first axis and the second axis intersect at a convergence point located within the processing chip.

57. The method according to claim 56, wherein the first axis and the second axis together define an inclination angle.

58. The method according to claim 57, wherein the inclination angle is in the range of about 10 degrees to about 45 degrees.

59. A method, the method comprising: conveying fluid through a mixing component of a processing chip to generate a mixture containing encapsulated particles in the fluid; monitoring the pressure of the fluid conveyed through the mixing component; activating a dynamic light scattering component to determine the particle size or particle size distribution of the encapsulated particles in the fluid, and when the fluid is in the processing chip, the dynamic light scattering component causes light to scatter off the particles; and Correlate the monitored pressure of the fluid with the determined particle size or particle size distribution. Transmitting the fluid through the mixing assembly includes transmitting a first fluid from a fluid input manifold channel to a first mixing assembly of a plurality of mixing assemblies, the fluid input manifold channel being fluidly coupled to a second mixing assembly of the plurality of mixing assemblies.

60. The method according to claim 59, further comprising using at least the monitored pressure to regulate the transmission of the fluid through the mixing assembly.

61. The method according to any one of claims 59 to 60, further comprising determining that the monitored pressure falls outside a predetermined range and, in response to determining that the monitored pressure falls outside the predetermined range, activating the dynamic light scattering assembly.

62. The method according to any one of claims 59 to 61, wherein activating the dynamic light scattering assembly comprises: Emitting light towards the encapsulated particles via a first optical fiber, the encapsulated particles scattering the emitted light, and the emitted light being transmitted through a light-transmissive material on a first side of the processing chip. Receiving the light scattered from the encapsulated particles, the received light being transmitted through the light-transmissive material on the first side of the processing chip, and the received light being received by a second optical fiber oriented obliquely with respect to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body located near the processing chip. Performing autocorrelation on the received light, and Using at least the autocorrelation to determine the particle size or particle size distribution of the encapsulated particles.

63. A method, the method comprises: Transmitting a fluid from a fluid input manifold channel of a processing chip through a first mixing assembly of the processing chip to generate a first mixture containing encapsulated particles in the fluid. Transmitting the fluid from the fluid input manifold channel through a second mixing assembly of the processing chip to generate a second mixture containing encapsulated particles in the fluid. Emitting light towards the encapsulated particles in the first mixture, the particles in the first mixture scattering the emitted light. Receiving the light scattered from the encapsulated particles in the first mixture. Performing autocorrelation on the received light scattered from the encapsulated particles in the first mixture. Using at least the autocorrelation of the received light scattered from the encapsulated particles in the first mixture to determine the particle size or particle size distribution of the encapsulated particles in the first mixture. Emitting light towards the encapsulated particles in the second mixture, the particles in the second mixture scattering the emitted light. Receiving the light scattered from the encapsulated particles in the second mixture. Performing autocorrelation on the received light scattered from the encapsulated particles in the second mixture. and Using at least the autocorrelation of the received light scattered from the encapsulated particles in the second mixture to determine the particle size or particle size distribution of the encapsulated particles in the second mixture.

64. The method according to claim 63, wherein emitting light towards the encapsulated particles in the first mixture, receiving light scattered from the encapsulated particles in the first mixture, emitting light towards the encapsulated particles in the second mixture, and receiving light scattered from the encapsulated particles in the second mixture are performed by a single dynamic light scattering assembly.

65. The method according to claim 64, further comprising: selectively delivering the first mixture to a fluid chamber of the single dynamic light scattering assembly, and emitting light towards the encapsulated particles in the first mixture and receiving light scattered from the encapsulated particles in the first mixture are performed when the first mixture is in the fluid chamber; and selectively delivering the second mixture to the fluid chamber, and emitting light towards the encapsulated particles in the second mixture and receiving light scattered from the encapsulated particles in the second mixture are performed when the second mixture is in the fluid chamber.

66. The method according to any one of claims 63 to 64, further comprising: activating a first dynamic light scattering assembly to emit light towards the encapsulated particles in the first mixture and receive light scattered from the encapsulated particles in the first mixture; and activating a second dynamic light scattering assembly, separate from the first dynamic light scattering assembly, to emit light towards the encapsulated particles in the second mixture and receive light scattered from the encapsulated particles in the second mixture.

67. The method according to claim 66, further comprising: selectively delivering the first mixture to a fluid chamber of a third dynamic light scattering assembly; emitting light towards the encapsulated particles in the first mixture when the first mixture is in the fluid chamber; receiving light scattered from the encapsulated particles in the first mixture when the first mixture is in the fluid chamber; selectively delivering the second mixture to the fluid chamber; emitting light towards the encapsulated particles in the second mixture when the second mixture is in the fluid chamber; and receiving light scattered from the encapsulated particles in the second mixture when the second mixture is in the fluid chamber.

68. The method according to claim 67, further comprising conveying the first mixture and the second mixture through a manifold, the first mixture and the second mixture passing through the manifold before reaching the fluid chamber.

69. An apparatus, comprising: a body including a first port and a second port, the body being positionable adjacent to a first outer surface of a processing chip; a first optical fiber coupled to the first port of the body, the first optical fiber for emitting light, the first port for guiding light emitted by the first optical fiber through a light-transmissive material of the processing chip and into a fluid chamber of the processing chip; a focusing lens supported by the body, the focusing lens being positioned and configured to focus light emitted by the first optical fiber; A second optical fiber, the second optical fiber being coupled to the second port of the body, the second optical fiber at the second port being oriented obliquely with respect to the first optical fiber at the first port, the second optical fiber being configured to receive light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber; A filter, the filter being supported by the body, the filter being positioned and configured to filter light scattered by particles in the fluid in the fluid chamber; and A processing chip, the processing chip being removably positioned relative to the body, the processing chip including: A first outer surface, A second outer surface, A fluid chamber, the fluid chamber being positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet; A light-transmissive material, the light-transmissive material being positioned between the first outer surface and the fluid chamber; A plurality of mixing components; and A fluid input manifold channel, the fluid input manifold channel being fluidly coupled to each of the plurality of mixing components.

70. The apparatus according to claim 69, further comprising a base having a processing chip mount for removably receiving the processing chip, the body being positioned adjacent to the processing chip mount.

71. The apparatus according to any one of claims 69 to 70, wherein the body is configured to orient the first optical fiber along an axis perpendicular to the outer surface of the processing chip.

72. The apparatus according to any one of claims 69 to 71, wherein the body is configured to orient the second optical fiber along an axis that is inclined with respect to the outer surface of the processing chip.

73. The apparatus according to any one of claims 69 to 72, further comprising a processor configured to receive data indicative of the light received by the second optical fiber and selectively direct fluid from the fluid input manifold channel to one of the plurality of mixing components based on the data.

74. An apparatus, comprising: A processing chip mount for removably receiving a processing chip; A body, the body including a first port and a second port, the body being fixedly secured relative to the processing chip mount, the processing chip mount being configured to removably receive the processing chip between the body and the processing chip mount; A first optical fiber, the first optical fiber being coupled to the first port of the body, the first optical fiber being configured to emit light, the first port being configured to direct the light emitted by the first optical fiber through a light-transmissive material of a processing chip received by the processing chip mount and into the fluid chamber of the processing chip; A second optical fiber, the second optical fiber being coupled to the second port of the body, the second optical fiber at the second port being oriented obliquely with respect to the first optical fiber at the first port, the second optical fiber being configured to receive light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber; and A processing chip removably received in the processing chip mount, the processing chip comprising: A first outer surface, A second outer surface, A fluid chamber positioned between the first outer surface and the second outer surface, the fluid chamber including a fluid chamber inlet and a fluid chamber outlet, A light-transmissive material positioned between the first outer surface and the fluid chamber, and A plurality of mixing components, and A fluid input manifold channel fluidly coupled to each of the plurality of mixing components.

75. The apparatus according to claim 74, wherein the processing chip is configured to form a therapeutic composition.

76. The apparatus according to claim 75, wherein the therapeutic composition comprises a fluid containing particles.

77. The apparatus according to claim 76, wherein the first port is for guiding light emitted by the first optical fiber through the light-transmissive material of the processing chip to the fluid of the therapeutic composition.

78. The apparatus according to claim 77, wherein the second optical fiber is for receiving light scattered by the particles of the therapeutic composition.

79. The apparatus according to claim 78, further comprising a processor to determine one or both of the following: determining the particle size of the particles in the therapeutic composition using at least the light scattered by the particles of the therapeutic composition or determining the particle size distribution of the particles in the therapeutic composition using at least the light scattered by the particles of the therapeutic composition.

80. The apparatus according to claim 79, wherein the processor is configured to selectively direct fluid from the fluid input manifold channel to one of the plurality of mixing components based on the determined one or both of the particle size of the particles in the therapeutic composition or the particle size distribution of the particles in the therapeutic composition.

81. An apparatus, Comprising: A processing chip, the processing chip comprising: A fluid chamber including a fluid chamber inlet and a fluid chamber outlet, and A light-transmissive material adjacent to the fluid chamber; A dynamic light scattering assembly, the processing chip being removably positioned relative to the dynamic light scattering assembly, the dynamic light scattering assembly for guiding light through the light-transmissive material and into the fluid chamber, the dynamic light scattering assembly further for receiving light scattered by particles in the fluid in the fluid chamber in response to the first optical fiber emitting light into the fluid chamber, and thereby capturing light scattering data; and A processor for determining the viscosity of the fluid in the fluid chamber based on the captured light scattering data, the processor further for determining one or both of the particle size or particle size distribution of the particles in the fluid based on the captured light scattering data, The processing chip further includes a plurality of mixing components, each of the plurality of mixing components having a plurality of inlets and an outlet, each of the plurality of mixing components being configured to form a mixture of fluids from the plurality of inlets and convey the mixture through the outlet, The processing chip further includes a fluid input port and a fluid input manifold channel, and the plurality of inlets of the plurality of mixing components are fluidly coupled to the fluid input manifold channel.

82. The apparatus according to claim 81, wherein the processing chip further comprises: a first channel, the fluid chamber inlet being configured to receive a first fluid from the first channel, and a second channel, the fluid chamber inlet further being configured to receive a second fluid from the second channel.

83. The apparatus according to claim 82, wherein the first fluid comprises a therapeutic composition.

84. The apparatus according to claim 83, wherein the therapeutic composition comprises at least some of the particles.

85. The apparatus according to claim 84, wherein the particles of the therapeutic composition comprise mRNA.

86. The apparatus according to any one of claims 82 to 85, wherein the second fluid comprises at least some of the particles.

87. The apparatus according to claim 86, wherein the particles of the second fluid comprise beads.

88. The apparatus according to claim 87, wherein the first fluid comprises a therapeutic composition, and the therapeutic composition comprises particles.

89. The apparatus according to claim 88, wherein the particles of the therapeutic composition have a first diameter, and the beads have a second diameter different from the first diameter.

90. The apparatus according to claim 89, wherein the second diameter is greater than the first diameter.

91. The apparatus according to any one of claims 82 to 90, wherein the first fluid comprises a first type of particles, and the second fluid comprises a second type of particles.

92. The apparatus according to claim 91, wherein the processor is configured to determine one or both of the particle size or the particle size distribution of the first type of particles in the first fluid based on light scattered by the first type of particles and the second type of particles.

93. The apparatus according to claim 92, wherein the second type of particles has a known particle size.

94. The apparatus according to any one of claims 82 to 93, wherein the second fluid comprises a diluent.

95. The apparatus according to claim 94, wherein the processing chip is configured to selectively add discrete amounts of the diluent to the first fluid in a sequence.

96. The apparatus according to claim 95, wherein the processing chip includes at least one valve to selectively control the delivery of the diluent to the first fluid.

97. The apparatus according to any one of claims 95 to 96, wherein the processing chip further includes at least one pump to selectively drive the movement of the diluent.

98. The apparatus according to any one of claims 94 to 97, wherein the processor is configured to: track the autocorrelation of the captured light scattering data during the sequence of adding discrete amounts of the diluent to the first fluid, and determine one or both of the particle size or the particle size distribution of the particles in the fluid based on the tracked autocorrelation.

99. The apparatus according to any one of claims 94 to 98, wherein the processing chip further comprises a mixing chamber to mix the diluent with the first fluid.

100. The apparatus according to claim 99, wherein the mixing chamber is positioned adjacent to the fluid chamber.

101. The apparatus according to any one of claims 99 to 100, wherein the processing chip further comprises a first pump and a second pump, the first pump and the second pump being configured to be alternately activated so as to drive the combination of the diluent and the first fluid back and forth through the mixing chamber.

102. A method, the method comprising: transmitting a fluid mixture through a processing chip, the fluid mixture containing particles; emitting light towards the fluid mixture via a first optical fiber, the particles in the fluid mixture scattering the emitted light; receiving the light scattered from the particles in the fluid mixture, the received light being received by a second optical fiber that is oriented obliquely with respect to the first optical fiber, the first optical fiber and the second optical fiber being fixed to a body located near the processing chip; performing autocorrelation on the received light; using at least the autocorrelation to determine the viscosity of the fluid mixture; and determining any one of the following: using at least the autocorrelation to determine the particle size of the particles in the fluid mixture, using at least the autocorrelation to determine the particle size distribution of the particles in the fluid mixture, or using at least the autocorrelation to determine the particle size and particle size distribution of the particles in the fluid mixture: Transmitting the fluid through the processing chip comprises: transmitting a first fluid component from a fluid input manifold channel to a first mixing assembly, the fluid input manifold channel being fluidly coupled to a second mixing assembly, the first fluid component including at least some of the particles; transmitting a second fluid component to the first mixing assembly, and mixing the first fluid component and the second fluid component together to form the fluid mixture.

103. The method according to claim 102, wherein the particles of the first fluid component include therapeutic particles.

104. The method according to claim 103, wherein the therapeutic particles include mRNA.

105. The method according to claim 104, wherein the mRNA is encapsulated in a delivery vehicle.

106. The method according to any one of claims 102 to 105, wherein the second fluid component includes at least some of the particles.

107. The method according to claim 106, wherein the particles of the second fluid component include beads.

108. The method according to any one of claims 106 to 107, wherein the particles of the first fluid component have a first diameter and the particles of the second fluid component have a second diameter different from the first diameter.

109. The method according to any one of claims 108, wherein the second diameter is greater than the first diameter.

110. The method according to any one of claims 106 to 109, wherein the particles of the first fluid component comprise particles of a first type, and the particles of the second fluid component comprise particles of a second type different from the particles of the first type.

111. The method according to any one of claims 106 to 110, receiving light scattered from the particles in the fluid mixture comprising: receiving light scattered by the particles of the first fluid component, and receiving light scattered by the particles of the second fluid component.

112. The method according to claim 111, determining any one of the following: determining the particle size of the particles in the fluid mixture using at least the autocorrelation, determining the particle size distribution of the particles in the fluid mixture using at least the autocorrelation, or determining the particle size and particle size distribution of the particles in the fluid mixture using at least the autocorrelation, including determining any one of the following: determining the particle size of the particles of the first fluid component using at least the autocorrelation, determining the particle size distribution of the particles of the first fluid component using at least the autocorrelation, or determining the particle size and particle size distribution of the particles of the first fluid component using at least the autocorrelation.

113. The method according to any one of claims 102 to 112, wherein the second fluid component comprises a diluent.

114. The method according to claim 113, conveying the fluid mixture through the processing chip further comprises adding discrete amounts of the diluent to the first fluid component in sequence.

115. The method according to claim 114, further comprising repeating emitting light, receiving light, and performing autocorrelation each time a discrete amount of the diluent is added to the first fluid component in the sequence.

116. The method according to claim 115, further comprising tracking the autocorrelation throughout each repetition of emitting light, receiving light, and performing autocorrelation each time a discrete amount of the diluent is added to the first fluid component in the sequence.

117. The method according to claim 116, determining the viscosity of the fluid mixture using at least the autocorrelation includes using the tracked autocorrelation to determine the viscosity of the fluid mixture.

118. The method according to any one of claims 116 to 117, determining any one of the following: determining the particle size of the particles in the fluid mixture using at least the autocorrelation, determining the particle size distribution of the particles in the fluid mixture using at least the autocorrelation, or determining the particle size and particle size distribution of the particles in the fluid mixture using at least the autocorrelation, including determining any one of the following: determining the particle size of the particles in the fluid mixture using the tracked autocorrelation, determining the particle size distribution of the particles in the fluid mixture using the tracked autocorrelation, or determining the particle size and particle size distribution of the particles in the fluid mixture using the tracked autocorrelation.

119. The method according to any one of claims 114 to 118, wherein the discrete amount of diluent added to the first fluid component is the same amount of diluent each time the discrete amount of diluent is added to the first fluid component according to the sequence.

120. The method according to any one of claims 102 to 119, wherein mixing the first fluid component and the second fluid component to form the fluid mixture includes alternately activating at least two pumps to drive the first fluid component and the second fluid component back and forth through a mixing chamber of the processing chip.

121. An apparatus comprising: a processing chip, the processing chip comprising: a first fluid input port, a first fluid input manifold fluidly coupled to the first fluid input port, a second fluid input port, a second fluid input manifold fluidly coupled to the second fluid input port, a plurality of mixing assemblies, each mixing assembly comprising: a first valve fluidly coupled to the first input manifold, a first inlet fluidly coupled to the first valve, a second valve fluidly coupled to the second input manifold, and a second inlet fluidly coupled to the second valve, an outlet, the mixing assembly being configured to form a mixture of fluids from the first inlet and the second inlet and convey the mixture out through the outlet; one or more measurement features, the one or more measurement features being operable to detect one or more characteristics of the mixture; and a processor configured to activate the first valve and the second valve of the plurality of mixing assemblies based on data from the one or more measurement features.

Images