Microfluidic device with elastic layer and curved surface

Abstract

The microfluidic device includes a first plate, a second plate, and a microfluidic path defined between the first and second plates. The microfluidic path includes at least one chamber. The microfluidic device also includes an elastic layer positioned between the first and second plates. The elastic layer includes a first membrane extending across at least one chamber. The first membrane is configured to deflect to drive fluid through at least one chamber. The elastic layer also includes a second membrane having at least one opening. At least one opening is aligned with at least one chamber.

Description

Technical Field

[0001] 〔Priority〕 This application claims the benefit of U.S. Patent Application No. 63 / 453,206, filed Mar. 20, 2023, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,” the disclosure of which is incorporated herein by reference.

Background Art

[0002] 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 discussed in this section, or problems associated with the subject matter provided as background, should not be assumed to have been recognized in the prior art. The subject matter of this section merely presents different approaches, which themselves may also be applicable to embodiments of the claimed technology.

[0003] 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 costly, too slow, or too prone to contamination for use in therapeutic formulations that may contain multiple polynucleotide species.

[0004] Details of one or more embodiments are set forth in the accompanying drawings and the description below. Other features, aspects, and advantages will be apparent from the description, drawings, and claims.

Brief Description of the Drawings

[0005] [Figure 1] Draw a schematic diagram of an example of a system including a microfluidic processing chip. [Figure 2] Draw an exploded perspective view of an example of the components of the system of FIG. 1. [Figure 3]Figure 1 shows a top view of one embodiment of a processing chip that can be incorporated into the system. [Figure 4A] A cross-sectional side view of the processing chip shown in Figure 3 is drawn in the first operating state. [Figure 4B] A cross-sectional side view of the processing chip shown in Figure 3, in the second operating state, is drawn. [Figure 4C] A cross-sectional side view of the processing chip shown in Figure 3, in the third operating state, is drawn. [Figure 4D] A cross-sectional side view of the processing chip shown in Figure 3, in the fourth operating state, is drawn. [Figure 4E] A cross-sectional side view of the processing chip shown in Figure 3 is drawn in the fifth operating state. [Figure 4F] A cross-sectional side view of the processing chip shown in Figure 3, in the sixth operating state, is drawn. [Figure 5] A top view of another embodiment of a processing chip that could be incorporated into the system shown in Figure 1 is drawn. [Figure 6] Figure 5 shows a disassembled and assembled perspective view of the processing chip, illustrating the elastic layers of the processing chip, including the upper and lower layers. [Figure 7] Figure 6 shows a top view of the lower membrane of the elastic layer. [Figure 8] Figure 6 shows a top view of the upper film of the elastic layer. [Figure 9A] Figure 5 shows a cross-sectional side view of the processing chip in the first operating state, illustrating the upper and lower regions of the valve chamber of the processing chip, which has a curved surface. [Figure 9B] Figure 5 shows a cross-sectional side view of the processing chip in the second operating state, and Figure 6 shows the lower film of the elastic layer that conforms to the curved surface of the lower region of the valve chamber. [Figure 9C] Figure 5 shows a cross-sectional side view of the processing chip in the third operating state, illustrating the lower film of the elastic layer in Figure 6, which conforms to the curved surface of the upper region of the valve chamber. [Figure 10] Figure 9A illustrates one example of a method for designing the valve chamber. [Figure 11] A cross-sectional side view of another embodiment of a processing chip that could be incorporated into the system shown in Figure 1 is drawn. [Modes for carrying out the invention]

[0006] In some embodiments, apparatus and methods for processing therapeutic polynucleotides are disclosed herein. In particular, these apparatus and methods may be closed-loop apparatus and methods configured to minimize or eliminate manual intervention during operation. Closed-loop apparatus and methods can provide a nearly completely sterile environment, and the components can provide a sterile pathway for processing from initial input (e.g., template) to output (e.g., formulated therapeutic agent). Material inputs to the apparatus (e.g., nucleotides and any chemical components) may be sterile and can be introduced into the system without requiring substantially any manual interaction.

[0007] The apparatus and methods described herein may be used to generate therapeutic agents with high reproducibility and rapid cycle times. The apparatus described herein may be configured to provide synthesis, purification, separation, compounding, and concentration of one or more therapeutic compositions in a single integrated apparatus. Alternatively, one or more of these processes may be carried out in two or more apparatuses as described herein. In some scenarios, the therapeutic composition may include therapeutic polynucleotides, such as ribonucleic acid or deoxyribonucleic acid. The polynucleotides may consist only of naturally occurring nucleotide units, or of any kind of synthetic, semi-synthetic, or modified nucleotide units. All or part of the processing steps may be carried out in an uninterrupted fluid processing pathway, which may be configured as one or a series of consumable microfluidic pathway devices, which may, in some cases herein, be referred to as a processing chip or biochip (although the chip does not necessarily have to be used in a bio-related application). In some embodiments, the processing chip may be removablely installed in an instrument that is part of a larger microfluidic system, such as the one shown in Figure 1. The disclosed apparatus and methods may be used for the synthesis of patient-specific therapeutic drugs, including compounding, in clinical settings (e.g., hospitals, clinics, pharmacies, etc.).

[0008] I. Terminology Throughout this specification and the subsequent claims, unless contextually required, the word “comprise,” and variations such as “comprises,” and “comprising,” mean that various components may be used together in a method and article (e.g., a composition, an apparatus including a device, a method). For example, the term “comprising” is understood to mean that it includes any described element or step, but does not exclude any other element or step. Generally, any apparatus and method described herein should be understood to be comprehensive, however all or a subset of components and / or steps may instead be exclusive and may be expressed as “consisting of” or instead “essentially consisting of” various components, steps, subcomponents, or substeps.

[0009] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural form unless otherwise indicated by the context. As used herein, the term “and / or” includes any combination of one or more of the related enumerated items and may be abbreviated as “ / .”

[0010] Spatial relative terms such as “under,” “below,” “lower,” “over,” and “upper” may be used herein to facilitate the description of the relationship between one element or feature and another, as shown in the drawings. It will be understood that spatial relative terms are intended to encompass different orientations of a device in use or operation, in addition to the orientation depicted in the drawings. For example, if a device in a drawing is inverted, an element described as being “beneath” or “below” another element or feature would be oriented “above” that other element or feature. Thus, the term “under” may encompass both upward and downward orientations. A device may be oriented differently (rotated 90° or oriented in another orientation), and the spatial relative descriptors used herein will be interpreted accordingly. Similarly, terms such as “upward,” “downward,” “vertical,” and “horizontal” are used herein for descriptive purposes only, unless otherwise indicated.

[0011] Where a feature or element is referred to herein as being "on" another feature or element, it may be directly above that other feature or element, or there may be intervening features and / or elements. Conversely, where a feature or element is referred to as being "directly above" another feature or element, there are no intervening features or elements. Where a feature or element is referred to as being "connected," "attached," or "linked" to another feature or element, it may be directly connected, attached, or linked to that other feature or element, or there may be intervening features or elements. Conversely, where a feature or element is referred to as being "directly connected," "directly attached," or "directly linked" to another feature or element, there are no intervening features or elements. Features and elements described or illustrated in this manner are described or illustrated in reference to one embodiment, but may apply to other embodiments as well. Furthermore, it will be understood by those skilled in the art that a reference to a structure or feature placed "adjacent" to another feature may have a portion that overlaps with or lies beneath that adjacent feature.

[0012] Where used herein and in the claims, including where used in the examples, and unless otherwise explicitly stated, all numbers can be read as if preceded by the words “about” or “approximately,” even if not explicitly stated. The words “about” or “approximately” can be used when describing magnitude and / or location to indicate that the described value and / or location is within a reasonable expected range of the value and / or location. For example, a number may have a value that is ±0.1% of the stated value (or range of values), ±1% of the stated value (or range of values), ±2% of the stated value (or range of values), ±5% of the stated value (or range of values), ±10% of the stated value (or range of values), and so on. Numbers described herein should also be understood to include about or approximately their value unless otherwise specified in the context. For example, if the value “10” is disclosed, “about 10” is also disclosed. Numerical ranges enumerated herein are intended to include all subranges contained within that range.

[0013] Furthermore, in a manner that can be appropriately understood by those skilled in the art, when a value is disclosed, it is understood that the ranges "less than or equal to" that value, "greater than or equal to" that value, and the possible ranges between values ​​are also disclosed. For example, if the value "X" is disclosed, then "less than or equal to X" and "greater than or equal to X" (for example, if X is a number) are also disclosed. It is also understood that throughout this application, data is provided in several different forms, and that this data represents a range of any combination of endpoints, starting points, and data points. For example, if a specific data point "10" and a specific data point "15" are disclosed, it is understood that "greater than 10 and 15", "greater than or equal to 10 and 15", "less than or equal to 10 and 15", and "the same as 10 and 15" are considered to be disclosed as well as the range between 10 and 15. It is also understood that each unit between two specific units is disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0014] The terms "first" and "second" may be used in this specification to describe various features / elements (including steps), but these features / elements should not be limited by these terms unless otherwise indicated in the context. These terms are used to distinguish one feature / element from another and do not indicate a particular order unless specifically stated. Thus, the first feature / element described hereinafter may be referred to as the second feature / element, and similarly, the second feature / element described hereinafter may be referred to as the first feature / element, without departing from the teachings of the present invention.

[0015] The terms "system", "apparatus", and "device" as used in this specification can be read as being interchangeable with each other. A system, apparatus, or device can each include a plurality of components having various kinds of structural and / or functional relationships with each other.

[0016] As used herein, "polynucleotide" refers to a nucleic acid molecule containing multiple nucleotides, and generally refers to both "oligonucleotides" (polynucleotide molecules with a length of 18 to 25 nucleotides) and polynucleotides with a length of 26 nucleotides or more. Aspects of this disclosure refer to oligonucleotides having a length of 18 to 25 nucleotides (e.g., 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, 24-mer, or 25-mer), or moderate-length polynucleotides having a length of 26 nucleotides or more (e.g., 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 ,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,approx.65,approx.70,approx.75,approx.80,approx.85,approx.90,approx.95,approx.100,approx.110,approx.120,approx.130,approx.140,approx.150,approx.160,approx.170,approx.180,approx.190,approx.200,approx.210,approx.220,approx.23 The composition comprises polynucleotides of 0, approximately 240, approximately 250, approximately 260, approximately 270, approximately 280, approximately 290, or approximately 300 nucleotides, or long polynucleotides having a length greater than approximately 300 nucleotides (e.g., approximately 300 to approximately 400 nucleotides, approximately 400 to approximately 500 nucleotides, approximately 500 to approximately 600 nucleotides, approximately 600 to approximately 700 nucleotides, approximately 700 to approximately 800 nucleotides, approximately 800 to approximately 900 nucleotides, approximately 900 to approximately 1000 nucleotides, approximately 300 to approximately 500 nucleotides, approximately 300 to approximately 600 nucleotides, approximately 300 to approximately 700 nucleotides, approximately 300 to approximately 800 nucleotides, approximately 300 to approximately 900 nucleotides, or polynucleotides of approximately 1000 nucleotides or longer). If the polynucleotide is double-stranded, its length can similarly be described in terms of base pairs.

[0017] As used herein, "amplification" can refer to the amplification of a polynucleotide. Amplification can include any suitable method for the amplification of a polynucleotide, including but not limited to multiple displacement amplification (MDA), polymerase chain reaction (PCR) amplification, loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification, strand displacement amplification, rolling circle amplification, and ligase chain reaction.

[0018] As used herein, a "cassette" (e.g., a synthetic in vitro transcription facilitator cassette) refers to a polynucleotide sequence that contains or can be operably linked to one or more expression elements such as an enhancer, promoter, leader, intron, 5' untranslated region (UTR), 3' UTR, or transcription termination sequence. In some embodiments, a cassette includes at least a first polynucleotide sequence capable of initiating transcription of an operably linked second polynucleotide sequence (which may include a template), and optionally, a transcription termination sequence operably linked to the second polynucleotide sequence. As described below, the template can include a sequence of interest, such as an open reading frame of interest (an "ORF"). A cassette can be provided as a single element or as two or more non-linked elements.

[0019] As used herein, "template" refers to a nucleic acid sequence that contains a sequence of interest for preparing a therapeutic polynucleotide according to the disclosed methods. The template can be, but is not limited to, double-stranded DNA (dsDNA), a modified plasmid construct, a cDNA sequence, or a linear nucleic acid sequence (e.g., a linear template generated by PCR or by annealing chemically synthesized oligonucleotides). In certain embodiments, the template can be incorporated into a "cassette" as described above.

[0020] As used herein, the term “sequence of interest” refers to a polynucleotide sequence that may be considered desirable for use for a suitable purpose, particularly for the production of mRNA for therapeutic purposes, and includes, but is not limited to, coding sequences of structural genes and non-coding regulatory sequences that do not code for mRNA or protein products.

[0021] As used herein, “in vitro transcription” or “IVT” refers to a process in which transcription is carried out in vitro in a non-cellular system to produce synthetic RNA molecules (e.g., synthetic mRNA) for use in a variety of applications, including therapeutic delivery to a subject, as therapeutic polynucleotides that may be part of or used in the formation of therapeutic polynucleotide compositions, such as those described below. The produced therapeutic polynucleotides (e.g., synthetic RNA molecules (transcripts)) can be combined with a delivery medium to form therapeutic polynucleotide compositions. Synthetic transcripts include mRNA, antisense RNA molecules, shRNA, circular RNA molecules, ribozymes, etc. The IVT reaction can use a purified linear DNA template containing a promoter sequence and an open reading frame (ORF) sequence of the sequence of interest, a buffer system containing ribonucleotide triphosphates or modified ribonucleotide triphosphates, DTT and magnesium ions, and phage RNA polymerase.

[0022] As used herein, “therapeutic polynucleotide” means a polynucleotide (e.g., mRNA) that may be part of a therapeutic polynucleotide composition delivered to a subject for the purpose of treating, preventing, or improving or altering the subject’s health condition.

[0023] As used herein, “therapeutic polynucleotide composition” (or, for short, “therapeutic composition”) may refer to a composition comprising one or more therapeutic polynucleotides (e.g., mRNA) encapsulated by a delivery medium, which may be administered to a subject in need by any suitable route of administration, such as intratumoral injection or intramuscular injection. An example of a therapeutic polynucleotide composition is mRNA (therapeutic) nanoparticles containing at least one mRNA encapsulated by a delivery medium molecule. mRNA vaccines are an example of a therapeutic polynucleotide composition.

[0024] As used herein, “delivery medium” refers to any substance that at least partially facilitates the in vivo, in vitro, or extra vivo delivery of polynucleotides (e.g., therapeutic polynucleotides) to target cells or tissues (e.g., tumors). Calling something a delivery medium does not preclude the possibility that the delivery medium also has therapeutic effects. Some versions of delivery mediums may offer additional therapeutic effects. In some versions, the delivery medium may be a peptoid molecule, such as an aminolipidized peptoid molecule, which can be used to at least partially encapsulate mRNA. The term “DV” is also used herein as an abbreviation for “delivery medium.”

[0025] As used herein, "joining" refers to methods for joining one component to another, such as ligation, synthesis, primer extension, annealing, recombination, or hybridization.

[0026] As used herein, "purifying" refers to the physical and / or chemical separation of one component (e.g., particles) from other unwanted components (e.g., contaminants, fragments, etc.).

[0027] As used herein, the term “substantially absent” in reference to a given substance includes not containing 100% of the given substance, or containing less than about 1.0%, less than about 0.5%, or less than about 0.1% of the given substance.

[0028] II. System Overview Including Microfluidic Processing Chip Figure 1 illustrates embodiments of various components that may be incorporated into System (100). In this embodiment, System (100) includes a housing (103) surrounding a seating mount (115) that can detachably hold one or more microfluidic processing chips (111). In other words, System (100) includes a chip receiving component configured to detachably house the processing chip (111), and the processing chip (111) itself defines one or more microfluidic channels or fluid pathways. Components of System (100) that fluidically interact with the processing chip (111) (e.g., within the housing (103)) may include fluid channels or pathways that are not necessarily considered microfluidic (e.g., such fluid channels or pathways are larger than the microfluidic channels or fluid pathways within the processing chip (111)). In some versions, the processing chip (111) is supplied and used as a disposable device, while the rest of System (100) is reusable. The housing (103) can take the form of a chamber, enclosure, or the like, with an opening that can be closed (e.g., via a lid or door) and thereby seal the interior. The housing (103) can enclose a temperature control device and / or be configured to enclose a temperature-controlled environment (e.g., a cooling unit). The housing (103) can form a sterile barrier. In some modifications, the housing (103) can form a humidified or humidity-controlled environment. In addition, or instead, the system (100) can be housed in a cabinet (not shown). Such a cabinet can provide a temperature-controlled (e.g., cooled) environment. Such a cabinet can also provide air filtration and airflow control, which can facilitate keeping reagents at a desired temperature throughout the manufacturing process. Furthermore, such a cabinet can be equipped with a UV lamp for sterilizing the processing tip (111) and other components of the system (100). Other suitable features can be incorporated into the cabinet housing the system (100).

[0029] In some scenarios, the assembly formed by the housing (103) and the components of the system (100) located inside the housing (103) may be considered a “device,” except for the processing chip (111). Although the controller (121) and user interface (123) are shown in Figure 1 as being outside the housing (103), the controller (121) and user interface (123) may actually be located inside or on top of the housing (103) and thus may form part of the device. As will be described in more detail below, this device can detachably receive the processing chip (111) via a seating mount (115). When the processing chip (111) is seated in the seating mount (115), the device and the processing chip (111) work together to form the system (100). When the processing chip (111) is removed from the seating mount (115), the remaining part of the system (100) may be considered a “device.” The apparatus, system (100), and processing chip (111) may each be considered “apparatus.” Thus, the term “apparatus” may be interpreted to include the apparatus alone, the processing chip (111) alone, a combination of the apparatus and the processing chip (111), any other combination of components of system (100), or system (100) or any other permutation of its components.

[0030] The seating mount (115) may 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. Thus, the seating mount (115) can facilitate the holding of the processing chip (111) in the appropriate position and orientation relative to other components of the system (100). In this embodiment, the seating mount (115) is configured to hold the processing chip (111) in a horizontal orientation, so that the processing chip (111) is parallel to the ground.

[0031] In some variations, the thermal control device (113) is located adjacent to the seating mount (115) and can regulate the temperature of any processing chip (111) mounted on the seating mount (115). The thermal control device (113) may include thermoelectric components (e.g., a Peltier element) and / or one or more heat sinks to control the temperature of all or part of any processing chip (111) mounted on the seating mount (115). In some variations, multiple thermal control devices (113) may be included, for example, to regulate the temperature of different areas of one or more regions of the processing chip (111) separately. The thermal control device (113) may include one or more thermal sensors (e.g., thermocouples) that can be used for feedback control of the processing chip (111) and / or the thermal control device (113).

[0032] As shown in Figure 1, the fluid interface assembly (109) connects the processing chip (111) to the pressure source (117), thereby providing one or more pathways for positive or negative pressure fluid (e.g., gas) to be transmitted 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 illustrated, the system (100) may include two or more pressure sources (117). In some scenarios, pressure may be generated by one or more sources other than the pressure source (117). For example, one or more vials or other fluid sources in the reagent storage frame (107) may be pressurized. Furthermore, or instead, reactions and / or other processes carried out on the processing chip (111) may generate additional fluid pressure. In this embodiment, the fluid interface assembly (109) also connects the processing chip (111) to the reagent storage frame (107), thereby providing one or more paths for the transfer of liquid reagents and the like from the reagent storage frame (107) to one or more internal regions of the processing chip (111), as will be described in more detail below.

[0033] In some versions, a pressurized fluid (e.g., gas) from at least one pressure source (117) reaches a fluid interface assembly (109) via a reagent storage frame (107), and the reagent storage frame (107) includes one or more components interposed in the fluid path between the pressure source (117) and the fluid interface assembly (109). In some versions, one or more pressure sources (117) are directly connected to the fluid interface assembly, and a positively pressurized fluid (e.g., a positively pressurized gas) or a negatively pressurized fluid (e.g., suction, or other negatively pressurized gas) bypasses the reagent storage frame (107) to reach the fluid interface assembly (109). Whether or not the fluid interface assembly (109) intervenes in the fluid path between the pressure source (117) and the fluid interface assembly (109), the fluid interface assembly (109) may be detachably connected to the rest of the system (100), and at least a portion of the fluid interface assembly (109) may be removed for sterilization between uses. As will be described in more detail below, the pressure source (117) can selectively pressurize one or more chamber regions on the processing tip (111). In addition, or alternatively, the pressure source can also selectively pressurize one or more vials or other fluid storage containers held by the reagent storage frame (107).

[0034] The reagent storage frame (107) is configured to accommodate multiple fluid sample holders, each of which may hold a fluid vial configured to hold a reagent (e.g., nucleotide, solvent, water, etc.) to be delivered to a processing chip (111). In some versions, one or more fluid vials or other storage containers within the reagent storage frame (107) may be configured to receive products from inside the processing chip (111). Furthermore, or alternatively, a second processing chip (111) may receive products from inside the first processing chip (111), and one or more fluids may be transferred from one processing chip (111) to another. In some such scenarios, the first processing chip (111) may perform a first dedicated function (e.g., synthesis, etc.), and the second processing chip (111) may perform a second dedicated function (e.g., encapsulation, etc.). The reagent storage frame (107) of this embodiment includes a manifold configured to divide a plurality of pressure lines and / or one or more pressure sources (117) into a plurality of pressure lines that can be applied to a processing chip (111). Such pressure lines can be controlled independently or collectively (in subcombinations).

[0035] The fluid interface assembly (109) may include a plurality of fluid lines and / or pressure lines, each such line including a biased (e.g., spring-loaded) holder or end that drives each fluid and / or pressure line to the processing tip (111) individually and independently when the processing tip (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 connected to the fluid interface assembly (109). In some versions, each fluid line includes a flexible tube that connects the reagent storage frame (107) to the processing tip (111) via a connector (e.g., a ferrule) that locks a vial into the tube. In some versions, the ends of the fluid lines / pressure lines may be configured to seal in contact with the processing tip (111) (e.g., in a corresponding seal port formed on the processing tip (111)), as described later. In this embodiment, the connections between the pressure source (117) and the processing tip (111), and the connections between the vial in the reagent storage frame (107) and the processing tip (111) all form sealed and closed pathways, which are isolated when the processing tip (111) is seated in the seating mount (115). Such sealed and closed pathways may provide protection against contamination when processing therapeutic polynucleotides.

[0036] The vials in the reagent storage frame (107) may be pressurized (e.g., to pressures greater than 1013.25 hPa (1 atm), e.g., 2026.5 hPa (2 atm), 3039.75 hPa (3 atm), 5066.25 hPa (5 atm), or higher). In some versions, the vials may be pressurized by a pressure source (117). Thus, negative or positive pressure may be applied. For example, fluid vials may be pressurized to approximately 6.89 to approximately 137.9 kPa (approximately 1 to approximately 20 psig) (e.g., 34.47 kPa (5 psig), 68.95 kPa (10 psig), etc.). Alternatively, a vacuum (e.g., approximately -48.26 kPa (approximately -7 psig) or approximately 48.26 kPa (approximately 7 psia)) may be applied to draw the fluid back into the vial (e.g., a vial that functions as a storage container) at the end of the process. The fluid vial can be driven at a lower pressure than the air valve, as described later, thereby preventing or reducing leakage. In some variations, the pressure difference between the fluid and the air valve may be approximately 6.89 kPa to approximately 172.37 kPa (approximately 1 psi to approximately 25 psi) (e.g., approximately 20.68 kPa (approximately 3 psi), approximately 34.47 kPa (approximately 5 psi), 48.26 kPa (7 psi), 68.95 kPa (10 psi), 82.74 kPa (12 psi), 103.42 kPa (15 psi), 137.9 kPa (20 psi), etc.).

[0037] The system (100) of this embodiment further includes a magnetic field applicator (119) configured to generate a magnetic field in the area of ​​the processing chip (111). The magnetic field applicator (119) may include a movable head which is operable to move the magnetic field and thereby selectively separate products adhering to magnetic trapping beads in vials or other storage containers in the reagent storage frame (107).

[0038] The system (100) of this embodiment further includes one or more sensors (105). In some versions, such sensors (105) include one or more cameras and / or other types of optical sensors. Such sensors (105) can sense one or more of the following: barcodes, fluid levels in fluid vials held within a reagent storage frame (107), fluid movement in a processing chip (111) mounted within a seating mount (115), and / or other optically detectable conditions. In versions in which sensors (105) are used to sense barcodes, such barcodes may be contained on vials in the reagent storage frame (107), and sensors (105) may be used to identify vials within the reagent storage frame (107). In some versions, a single sensor (105) is positioned and configured to simultaneously visualize barcodes on vials in a reagent storage frame (107), fluid levels in vials in the reagent storage frame (107), fluid movement in a processing chip (111) mounted in a seating mount (115), and / or other optically detectable conditions. In some other versions, multiple sensors (105) are used to visualize such conditions. In some such versions, different sensors (105) may be positioned and configured to separately visualize corresponding optically detectable conditions, so that the sensors (105) can be dedicated to specific corresponding optically detectable conditions.

[0039] In versions of sensor (105) that include at least one optical sensor, visual / optical markers may be used to estimate yield. For example, fluorescence may be used to detect process yield or residue by labeling with a phosphor. In addition, or instead, dynamic light scattering (DLS) may be used to measure particle size distribution within a portion of the processing chip (111) (e.g., a mixed portion of the processing chip (111)). In some modifications, sensor (105) may provide measurements using one or two optical fibers to carry light (e.g., laser light) into the processing chip (111); and the optical signal exiting the processing chip (111) may be detected. In versions of sensor (105) that optically detect process yield or residue, etc., sensor (105) may be configured to detect visible light, fluorescence, ultraviolet (UV) absorption signals, infrared (IR) absorption signals, and / or any other suitable type of optical feedback.

[0040] In a version including at least one optical sensor (105) configured to capture video images, such sensor (105) can record at least some of the activity on the processing chip (111). For example, the entire process 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) that can visualize the processing chip (111) (e.g., from above). The processing on the processing chip (111) may be visually tracked, and this video recording may be retained for later quality control and / or processing. Thus, the video recording of this processing may be stored, stored, and / or transmitted for subsequent review and / or analysis. Furthermore, as will be described in more detail below, the video may be used as a real-time feedback input that can influence the processing using at least visually observable states captured in the video.

[0041] The system (100) of this embodiment may be controlled by a controller (121). The controller (121) may include one or more processors, one or more memories, and various other suitable electrical components. In some versions, one or more components of the controller (121) (e.g., one or more processors) are integrated into the system (100) (e.g., housed in a housing (103)). In addition, or instead, one or more components of the controller (121) (e.g., one or more processors) may be detachably attached to or detachably connected to other components of the system (100). Thus, at least a portion of the controller (121) can be detachable. Furthermore, in some versions, at least a portion of the controller (121) may be separate from the housing (103).

[0042] Control by the controller (121) may include, among several tasks, activating a pressure source (117) and applying pressure through a processing chip (111) to drive fluid movement. The controller (121) may be entirely or partially outside the housing (103), or entirely or partially inside the housing (103). The controller (121) may be configured to receive user input via the system (100)'s user interface (123) and to provide output to the user via the user interface (123). In some versions, the controller (121) is fully automated up to a stage where user input is not required. In some such versions, the user interface (123) may only provide output to the user. The user interface (123) may include a monitor, touchscreen, keyboard, and / or any other suitable features. The controller (121) can coordinate processes including moving one or more fluids onto and over 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 fluid in the processing chip (111), adjusting the temperature of the processing chip (111), and applying a magnetic field (for example, when using magnetic beads). The controller (121) can receive real-time feedback from the sensor (105) and execute a control algorithm in accordance with such feedback from the sensor (105). Such feedback from the sensor (105) may include, but are not limited to, the identification of reagents in vials in the reagent storage frame (107), the detected fluid level in vials in the reagent storage frame (107), the detected movement of fluid in the processing chip (111), and the fluorescence of phosphors in the fluid in the processing chip (111). The controller (121) may include software, firmware, and / or hardware.The controller (121) can also communicate with a remote server for purposes such as tracking the operation of the device, reordering materials (e.g., nucleotides, components such as the processing chip (111)), and / or downloading protocols.

[0043] Figure 2 shows an embodiment of a specific form that the various components of the system (100) can take. In particular, Figure 2 shows a reagent storage frame (150), a fluid interface assembly (152), a seating mount (154), a thermal control device (156), and a processing chip (200). The reagent storage frame (150), fluid interface assembly (152), seating mount (154), thermal control device (156), and processing chip (200) in this embodiment can be configured and operated in the same way as the reagent storage frame (107), fluid interface assembly (109), seating mount (115), thermal control device (113), and processing chip (111) described above. These components are fixed to a base (180). A pair of rods (182) support the reagent storage frame (150) on the fluid interface assembly (152).

[0044] As shown in Figure 2, a pair of optical sensors (160) are positioned at four separate locations along the base (180). The optical sensors (160) can be configured and operated in the same manner as the sensor (105) described above. The optical sensors (160) can include commercially available cameras or any other suitable type of optical sensor. The optical sensors (160) are positioned so that fluid vials held within the reagent storage frame (150) are within the field of view of one or more of the optical sensors (160). In addition, the 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 rail (184) (e.g., in a gantry configuration), and each optical sensor (160) is configured to translate laterally along its respective corresponding rail (184). Linear actuators (186) are fixed to each optical sensor (160) and are thereby operable to drive the lateral translational motion of each optical sensor (160) along the corresponding rail (184). Each actuator (186) may be in the form of a drive belt, drive chain, drive cable, or any other suitable type of structure. A controller (121) may drive the operation of the actuators (186). The optical sensors (160) may be moved along the rails (184) during the operation of the system (100) to facilitate visualization of suitable areas of vials and / or processing tips (200) within the reagent storage frame (150). In some scenarios, the optical sensors (160) may move simultaneously along the corresponding rails (184). In some other scenarios, the optical sensors (160) may move independently along the corresponding rails (184).

[0045] Although the optical sensor (160) is shown mounted on the base (180) in Figure 2, the optical sensor (160) may be positioned elsewhere in the system (100) in addition to, or instead of, being mounted on the base (180). For example, some versions of the reagent storage frame (107) may include one or more optical sensors (160) positioned and configured to provide a top-down view. In some such versions, the optical sensor (160) may be mounted on a rail, a movable cantilever, or other structure that allows the optical sensor (160) to change position during the operation of the system (100). The optical sensor (160) may be positioned in any other suitable location. Although not shown, the system (100) may also include one or more light sources (e.g., an electroluminescent panel) to provide illumination to assist optical sensing by the optical sensor (160).

[0046] In some versions, one or more mirrors may be used to facilitate the visualization of components of the system (100) by the optical sensor (160). Such mirrors allow the optical sensor (160) to see components of the system (100) that might not otherwise be within the field of view of the sensor (160). Such mirrors may be positioned directly adjacent to the optical sensor (160). In addition, or instead, such mirrors may be positioned adjacent to one or more components of the system (100) that are visible to the optical sensor (160).

[0047] While using the system (100), the operator can select a protocol to execute (for example, from a library of pre-configured protocols), or the user can input a new protocol (or modify an existing one) via the user interface (123). From this protocol, the controller (121) can instruct the operator on what type of processing chip (111) to use, what the contents of the vials in the reagent storage frame (107) should be, and where to place the vials in the reagent storage frame (107). The operator can load the processing chip (111) into the seating mount (115) and load the desired reagent vials and discharge vials into the reagent storage frame (107). The system (100) can verify the presence of the desired peripherals, identify the processing chip (111), and scan the identifier (e.g., barcode) of each reagent and product vial in the reagent storage frame (107) to facilitate matching the vials with the reagent table of the selected protocol. After confirming the initial materials and equipment, the controller (121) can execute the protocol. During execution, valves and pumps are activated to deliver reagents, the reagents are blended, the temperature is controlled, the reaction takes place, measurements are taken, and the product is delivered to the target vial in the reagent storage frame (107), as will be described in more detail below.

[0048] III. Examples of Processing Chips Figures 3 and 4A–4F illustrate an embodiment of the processing chip (200) in further detail. Combined with the rest of the system (100), the processing chip (200) can be used to provide in vitro synthesis, purification, concentration, preparation, and / or analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions. As shown in Figure 3, the processing chip (200) in this embodiment includes a plurality of fluid ports (220). Each fluid port (220) has an associated fluid channel (222) formed within the processing chip (200), and the fluid transmitted into the fluid port (220) flows through the corresponding fluid channel (222). As will be 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 embodiment, each fluid channel (222) leads to a valve chamber (224), which is operable to selectively prevent or allow the fluid from the corresponding fluid channel (222) to be further transmitted along the processing chip (200), as will be described in more detail below.

[0049] As shown in Figure 3, the processing tip (200) in this embodiment includes several additional chambers (230, 250, 270) which can be used to serve various purposes during the process of manufacturing therapeutic compositions as described herein. In just one example, such additional chambers (230, 250, 270) may be used to provide synthesis, purification, separation, compounding, and / or concentration of one or more therapeutic compositions, or to perform any other suitable function. Fluids may be transferred from one chamber (230) to another (230) via fluid connectors (232). In some versions, the fluid connectors (232) can operate like valves between open and closed states (similar to, for example, valve chambers (224)). In some other versions, the fluid connectors (232) remain open throughout the entire process of manufacturing therapeutic compositions. In this embodiment, chamber (230) is used to provide the synthesis of polynucleotides, but chamber (230) could also serve any other suitable purpose.

[0050] In the embodiment shown in Figure 3, another valve chamber (234) is placed between one of the chambers (230) and one of the chambers (250), thereby allowing the fluid to be selectively transferred from chamber (230) to chamber (250). The chambers (250) are provided in pairs and connected to one another, and the processing tip (200) can transfer the fluid back and forth between the chambers (250). In this embodiment, a pair of chambers (250) are provided, but any other suitable number of chambers (250), including just one chamber (250) or two or more chambers (250), may be used. The chambers (250) may be used to provide purification of a fluid and / or serve any of the various other purposes described herein and may have any suitable configuration. In the version in which the chambers (250) are used for purification, the chambers (250) may include a material configured to absorb a selected portion from the fluid mixture within the chambers (250). In some such versions, the material may include a cellulose material that can selectively absorb double-stranded mRNA from the mixture. In some such versions, the cellulose material may be inserted into only one of a pair of chambers (250), and when the fluid from the first chamber (250) of this pair is mixed into the second chamber (250), mRNA and / or some other components can be effectively removed from the fluid mixture, which can then be transferred to another pair of chambers (270) further downstream for further processing or discharge. Alternatively, the chambers (250) may be used for any other suitable purpose.

[0051] An additional valve chamber (252) is placed between each chamber (250) and the corresponding chamber (270), thereby allowing fluid to be selectively transferred from chamber (250) to chamber (270) via the valve chamber (252). The chambers (270) are also connected to one another, and the processing tip (200) can transfer fluid back and forth between the chambers (270). The chambers (270) may be used to provide a mixture of fluids and / or may serve any of the various other purposes described herein and may have any suitable configuration.

[0052] As shown in Figure 3, the chamber (270) is also connected to an additional fluid port (221) via a corresponding fluid channel (223) and valve chamber (225). The fluid port (221), fluid channel (223), and valve chamber (225) can be configured and made operational in the same way as the aforementioned fluid port (220), fluid channel (222), and valve chamber (224). In some versions, the fluid port (221) is used to deliver additional fluid to the chamber (270). In addition, or instead, the fluid port (221) may be used to deliver fluid from the processing chip (200) to another device. For example, fluid from the chamber (270) may be delivered directly via the fluid port (221) to another processing chip (200), to one or more vials in a reagent storage frame (107), or to another location.

[0053] The processing tip (200) further includes a plurality of reservoir chambers (260). In this embodiment, each reservoir chamber (260) is configured to receive and store fluid being transferred to or from the corresponding chambers (250, 270). Each reservoir chamber (260) has a corresponding inlet valve chamber (262) and an outlet valve chamber (264). Each inlet valve chamber (262) is positioned between the reservoir chamber (260) and the corresponding chambers (250, 270) and is operable to allow or prevent the flow of fluid between the reservoir chamber (260) and the corresponding chambers (250, 270). Each outlet valve chamber (264) is operable to meter the flow of fluid between the reservoir chamber (260) and the corresponding fluid port (266). In some versions, each fluid port (266) is configured to transfer fluid from the corresponding vial in the reagent storage frame (107) to the corresponding reservoir chamber (260). Furthermore, or alternatively, each fluid port (266) may be configured to transfer fluid from the corresponding reservoir chamber (260) to the corresponding vial in the reagent storage frame (107). In this embodiment, the reservoir chamber (260) is used to provide metering of the fluid transferred to and / or from the processing tip (200). Alternatively, the reservoir chamber (260) may be used for any other suitable purpose, including but not limited to pressurizing the fluid transferred to and / or from the processing tip (200).

[0054] As shown in Figure 3, the processing chip (200) in this embodiment includes a plurality of pressure ports (240). Each pressure port (240) has an associated pressure channel (244) formed within the processing chip (200), and the pressurized gas transmitted through the pressure port (240) is further transmitted through the corresponding pressure channel (244). As will be 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 embodiment, each pressure channel (244) leads to a corresponding chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270), thereby providing valve action or peristaltic pumping through the chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270), as will be described in more detail below.

[0055] The processing chip (200) may also 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 the embodiment shown in Figure 3, the processing chip (200) includes an electrically active region (212) containing such electrical communication features. The electrically active region (212) may further include electrical circuits and other electrical components. In some versions, the electrically active region (212) may provide transmission of power, data, etc. Although the electrically active region (212) is shown at one specific location on the processing chip, the electrically active region (212) may instead be located at one or more other suitable locations. In some versions, the electrically active region (212) is omitted.

[0056] As shown in Figures 4A to 4F, the processing chip (200) further includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306). As will be described in more detail below, some versions 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), with the lower surface (310) juxtaposed with the elastic layer (302). The second plate (304) has an upper surface (312) and a lower surface (314), with the upper surface (312) juxtaposed with the elastic layer (302) and the lower surface (314) juxtaposed with the third plate (306). Thus, the elastic layer (302) is placed between the first plate (300) and the second plate (304). In this embodiment, another elastic layer (316) is also placed between the second plate (304) and the third plate (306), although this elastic layer (316) is optional.

[0057] The plates (300, 304, 306) in this embodiment are substantially translucent to visible light and / or ultraviolet light. "Substantially translucent" means that at least 90% (and possibly 100%) of light is transmitted through the material compared to a translucent material. In some modifications, 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 transparent" means that at least 90% (and possibly 100%) of light is transmitted through the material compared to a completely transparent material. In another embodiment, one or more of the plates (300, 304, 306) may transmit ultraviolet light at a wavelength of about 260 nm with a transmittance ranging from about 0.2% to about 20%, including about 0.4% to about 15%, or about 0.5% to about 10%.

[0058] Furthermore, the plates (300, 304, 306) in this embodiment are rigid. In some other versions, one or more of the plates (300, 304, 306) are semi-rigid. The plates (300, 304, 306) may include glass, plastic, silicone, and / or any other suitable material. In some versions, one or more of the plates (300, 304, 306) are formed as a laminate of two or more layers of material, so that each plate (300, 304, 306) does not necessarily have to be formed as a single homogeneous continuum of material. The material constituting one of the plates (300, 304, 306) may be different from the material constituting the other plates (300, 304, 306).

[0059] In this embodiment, the elastic layer (302) is formed as a flexible film that is impermeable to liquids. In some versions, the elastic layer (302) is gas permeable despite being impermeable to liquids. In some such versions, certain areas of the elastic layer (302) are treated to be gas permeable, while untreated areas of the elastic layer (302) remain gas impermeable. As will be discussed later, the elastic layer (302) may be used to drive fluid across the processing tip (200) via peristaltic pump action. Also as will be discussed later, the elastic layer (302) may be used to provide valves at various locations along the processing tip (200). In some versions, a single sheet of elastic material spans the entire width of the processing tip (200) to form the elastic layer (302). In some other versions, two or more separate pieces of elastic material are used to form the elastic layer (302), and such separate pieces of elastic material are positioned at different locations across the width of the processing tip (200). As just one example, the elastic layer (302) may include a film containing a polydimethylsilicone (PDMS) elastomer film.

[0060] As is best seen in Figures 4A to 4F, the first plate (300) and the second plate (304) work together to define multiple chambers (320, 322, 324, 326), and the elastic layer (302) divides each chamber (320, 322, 324, 326) into a corresponding upper chamber region (330) and a lower chamber region (332). The chambers shown in Figure 3 (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) can be configured and operated in the same way as the chambers shown in Figures 4A to 4F (320, 322, 324, 326). For example, chamber (320) may be similar to chamber (264), chamber (322) may be similar to chamber (260), chamber (324) may be similar to chamber (262), and chamber (326) may be similar to chamber (250).

[0061] As shown in Figures 4A to 4F, the fluid port (220) is formed through the first plate (300). A corresponding opening (342) is formed through the region of the elastic layer (302) below the fluid port (220). The fluid channel (222) extends from the opening (342) to the lower chamber region (332) of the first chamber (320). As previously stated, the fluid port (220) is configured to receive the fluid line (206) from the fluid interface assembly (109). The distal end of the fluid line (206) is configured to seal in contact with the region of the elastic layer (302) exposed by the fluid port (220) and to transmit the fluid (207) through the opening (342). In some versions, a spring or other elastic member provides elastic bias to the fluid line (206), pressing the distal end of the fluid line (206) against the region of the elastic layer (302) exposed by the fluid port (220), thereby maintaining the seal. The fluid (207) from the fluid line (206) reaches the lower chamber region (332) of the first chamber (320) via the fluid channel (222). As will be described in more detail below, this fluid (207) may be further transferred from the first chamber (320) to other chambers (322, 324, 326) by peristaltic pumping action provided through the elastic layer (302). After reaching the fourth chamber (326), the fluid (207) may be further transferred to other chambers or other features in the processing tip (200), to storage vials in the reagent storage frame (107), or otherwise processed. Therefore, the fluid (207) path does not necessarily terminate in the fourth chamber (326). Furthermore, it should be understood that the other fluid ports (221, 266) shown in Figure 3 can all be configured and operated in the same way as the fluid port (220) shown in Figures 4A to 4F.

[0062] The pressure port (240) is formed through the first plate (300). A corresponding opening (344) is formed through the region of the elastic layer (302) below the pressure port (240). The pressure channel (244) extends from the opening (344) to the upper chamber region (330) of the first chamber (320). As previously stated, the pressure port (240) is configured to receive a pressure line (208) from the fluid interface assembly (109) and thereby receive pressurized gas from the pressure source (117). The distal end of the pressure line (208) is sealed in contact with the region of the elastic layer (302) exposed by the pressure port (240) and is configured to transmit positively or negatively pressurized gas through the opening (344). In some versions, a spring or other elastic member provides elastic bias to the pressure line (208), pressing the distal end of the pressure line (208) against the region of the elastic layer (302) exposed by the pressure port (240), thereby maintaining the seal. The positively or negatively pressurized gas from the pressure line (208) reaches the upper chamber region (330) of the fourth chamber (326) via the pressure channel (244).

[0063] Figures 4A to 4F depict a configuration where only one pressure line (208) is connected to the processing chip (200). However, the processing chip (200) may have multiple connected pressure lines (208), each independently applying positive or negative pressure to the corresponding chambers (320, 322, 324, 326) of the processing chip (200). In some versions, one or more of the chambers (320, 322, 324, 326) have their own dedicated pressure line (208) and corresponding pressure channel (244). Furthermore, 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). Figures 4A to 4F illustrate pressure channels (244) formed through the second plate (304), however some pressure channels (244) (or regions of pressure channels (244)) may be formed by the first plate (300). For example, some pressure channels (244) (or regions of pressure channels (244)) may be formed between a recess on the lower surface of the first plate (300) and the upper surface of the elastic layer (302).

[0064] IV. Examples of valve action and peristaltic pump action driven via an elastic layer As described above, the elastic layer (302) can operate to drive the fluid through the processing tip (200) by peristaltic pump action and to stop the movement of the fluid through the processing tip (200) by providing valve action. An example of such operation is illustrated in the sequence drawn across Figures 4A to 4F. In this embodiment, chambers (320, 324) function as valve chambers, and chamber (322) function as a metering chamber. Chamber (326) functions as a working chamber, where synthesis, purification, separation, compounding, concentration, or any other process is performed. This configuration, arrangement, and use of chambers (320, 322, 324, 326) is provided as an exemplary embodiment. Chambers (320, 322, 324, 326) may be configured, arranged, and used in other ways instead.

[0065] Figure 4A shows the processing tip (200) before the fluid and pressurized gas have been transferred to it. In Figure 4B, the positively pressurized gas has been transferred to the upper chamber region (330) of the chamber (324), the negatively pressurized gas has been transferred to the upper region (330) of the chambers (320, 322), and the fluid (207) has been transferred to the chambers (320, 322). In this state, the positively pressurized gas deforms the elastic layer (302) inside the chamber (324), and the elastic layer (302) sits in contact with the surface of the lower chamber region (332) of the chamber (324). The elastic layer (302) seats in contact with the surface of the lower chamber region (332) of the chamber (324), preventing the fluid (207) from entering the chamber (324), thereby causing the chamber (324) to function like a shut-off valve in the state shown in Figure 4B. The negative pressure exerted by the gas in the upper chamber region (330) of the chambers (320, 322) deforms the corresponding portion of the elastic layer (302) inside the chambers (320, 322), causing it to seat in contact with the upper chamber region (330) of the chambers (320, 322). This allows the fluid (207) to occupy the entire volume of the chambers (320, 322).

[0066] After reaching the state shown in Figure 4B, the positively pressurized gas is transferred to the upper chamber region (330) of the chamber (320), but the air pressure in the chambers (322, 324) can remain unchanged. This results in the state shown in Figure 4C. As shown, the positively pressurized gas deforms the elastic layer (302) inside the chamber (320), causing the elastic layer (302) to seat in contact with the surface of the lower chamber region (332) of the chamber (320). The seating of the elastic layer (302) in contact with the surface of the lower chamber region (332) of the chamber (320) drives the fluid (207) out of the chamber (320), causing the chamber (320) to act like a shut-off valve in the state shown in Figure 4C. However, the volume of the fluid (207) inside the chamber (322) is not affected in the state shown in Figure 4C. Therefore, the chamber (322) can be used to provide metering of the fluid (207), thereby ensuring that only a precise, predetermined volume of fluid (207) is further transferred along the processing tip (200). As just one example, such metering volumes can be approximately 10 nL, 20 nL, 25 nL, 50 nL, 75 nL, 100 nL, 1 microliter, 5 microliters, etc.

[0067] Once the appropriate metering volume is achieved, the negatively pressurized gas is transferred to the upper chamber region (330) of the chambers (324, 326), and the air pressure state in the chambers (320, 322) can remain unchanged. This results in the state shown in Figure 4D. As shown in the figure, the negatively pressurized gas in the upper chamber region (330) of the chambers (324, 326) deforms the corresponding portion of the elastic layer (302) in the chambers (324, 326), causing it to seat in contact with the surface of the upper chamber region (330) of the chambers (324, 326). This effectively opens the valve formed by the chamber (324), and the chamber (326) becomes ready to receive the fluid (207). This also generates negative pressure in the chamber (324), which draws the fluid (207) from the chamber (322) into the chamber (324).

[0068] While the valve formed by chamber (324) is open, the positively pressurized gas is transmitted to the upper chamber region (330) of chamber (322), and the pneumatic state of chambers (320, 324, 326) can remain unchanged. This results in the state shown in Figure 4E. As shown, the positively pressurized gas in the upper chamber region (330) of chamber (322) deforms the corresponding portion of the elastic layer (302) in chamber (322), causing it to seat in contact with the surface of the lower chamber region (332) of chamber (322). This deformation of the elastic layer (302) drives the fluid (207) out of chamber (322). Since the valve formed by chamber (320) is closed and the valve formed by chamber (324) is open, the fluid (207) moves from chamber (322) into chamber (324). In this embodiment, since the volume of chamber (322) is larger than the volume of chamber (324), the fluid (207) from chamber (322) overflows from chamber (324) into chamber (326).

[0069] When the fluid (207) is transferred from chamber (322) to chambers (324, 326), the positively pressurized gas is transferred to the upper chamber region (330) of chamber (324), and the air pressure state in chambers (320, 322, 326) can be kept unchanged. This results in the state shown in Figure 4F. As shown in the figure, the positively pressurized gas in the upper chamber region (330) of chamber (324) deforms the corresponding portion of the elastic layer (302) in chamber (324), causing it to seat in contact with the surface of the lower chamber region (332) of chamber (324). This deformation of the elastic layer (302) drives the fluid (207) out of chamber (324). The deformed portion of the elastic layer (302) within the chamber (324) effectively seals the chamber (324) away from the chamber (322) (for example, so that the chamber (324) acts like a valve in a closed state), and the fluid (207) moves from the chamber (324) into the chamber (326).

[0070] At the stage shown in Figure 4F, the fluid (207) has been discharged from the chambers (320, 332, 324), and chamber (326) contains a precisely measured volume of fluid (207) from chamber (322). The fluid (207) in chamber (326) may be further processed within chamber (326) in accordance with the teachings herein. In addition, or instead, the fluid (207) in chamber (326) may be transferred to one or more other chambers in the processing tip (200), or to vials in the reagent storage frame (107), or otherwise handled. Regardless of how the fluid (207) is handled after it reaches the chamber (326), it should be understood that the fluid (207) is transported along the chambers (320, 322, 324) in a certain order so that it reaches the chamber (326) via peristaltic action produced by the elastic layer (302) in response to the transport of positively or negatively pressurized gases to the upper chamber region (330) of the chambers (320, 322, 324, 326) in a specific order. Such peristaltic pumping action may have special advantages for moving fluids that may be viscous or contain suspended particles such as purifying beads or trapping beads. Such peristaltic pumping action by selective deformation of the elastic layer (302) may also be called pneumatic barrier deflection or "pneumodeflection".

[0071] In some scenarios, it may be desirable to remove air or other gases from one or more fluid pathways within the processing chip (200). To achieve this, the processing chip (200) may include one or more chambers configured to provide ventilation to the fluid pathways or, otherwise, to vent gases from the fluid pathways. For example, such ventilation or venting may be performed as part of a priming process when the fluid is first introduced into the processing chip (200). Alternatively, such ventilation or venting may be performed to remove gases generated in the fluid during the process of forming a therapeutic composition. Such ventilation or gas removal chambers may be called “vacuum caps”. In some versions, at least a region of the elastic layer (302) located within the vacuum cap (if not the entire elastic layer (302)) is gas permeable (while still liquid impermeable). A negatively pressurized gas may be applied to an upper chamber region (330) of the chamber used as a vacuum cap, and this negatively pressurized gas can draw air or gases from the fluid pathways through the corresponding region of the elastic layer (302). In some versions, the upper chamber region (330) of the chamber used as a vacuum cap includes one or more protrusions or standoff features, which prevent the corresponding region of the elastic layer (302) from contacting and fully seating on the surface of the upper chamber region (330) of the chamber used as a vacuum cap. This can further facilitate the discharge of air or other gases through the vacuum cap.

[0072] V. Example of a processing chip having improved pumping / valving characteristics As described above, the elastic layer (302) is placed between the first plate (300) and the second plate (304) and may be used to drive fluid across the processing tip (200) via peristaltic pump action and / or to provide valves at various locations along the processing tip (200). In some scenarios, it may be desirable to provide an elastic layer (302) with increased thickness, such as within the area of ​​the elastic layer (302) bonded to the first plate (300) and / or the second plate (304). For example, increasing the thickness of the elastic layer (302) in such area can improve the seal between that area of ​​the elastic layer (302) and one or both of the first plate (300) and / or the second plate (304). In other words, the desire to improve the effectiveness of the seal at the interface between the elastic layer (302) and the plates (300, 304) may drive the tendency to increase the thickness of the elastic layer (302). Furthermore, or alternatively, by increasing the thickness of the elastic layer (302), the diffusion of air (or other gases) can be reduced in situations where such diffusion is unnecessary or otherwise undesirable, such as when the valve is closed.

[0073] Furthermore, it may be desirable to provide an elastic layer (302) with minimal thickness in areas of the elastic layer (302) located inside any of the chambers (320, 322, 324, 326). For example, reducing the thickness of the elastic layer (302) in such areas may improve the response of that area of ​​the elastic layer (302) to pneumatic pressure, by allowing that area of ​​the elastic layer (302) to conform more closely to the surface of the upper chamber region (330) and / or lower chamber region (332) of each chamber (320, 322, 324, 326) in response to pneumatic pressure (supplied, for example, via the pressure line (208)), thereby enabling it to contact and seat more completely on that surface. In other words, the desire to enhance the effectiveness of valve action and peristaltic pump action through deformation of the elastic layer (302) may encourage a tendency to reduce the thickness of the elastic layer (302). Furthermore, or alternatively, reducing the thickness of the elastic layer (302) in the region may improve the diffusion of air (or other gases) in situations where such diffusion is necessary or otherwise desirable, for example, during priming when the fluid is first introduced into the processing tip (200) and / or during the discharge of air or other gases through the vacuum cap. This tendency to reduce the thickness of the elastic layer (302) (to enhance the effectiveness of valve action and peristaltic pump action) can often conflict with the aforementioned tendency to increase the thickness of the elastic layer (302) (to strengthen the seal at the interface). Therefore, these conflicting demands and trends may require a compromise that effectively achieves both demands simultaneously. Examples of how such a compromise may be achieved are described in more detail below.

[0074] Furthermore, or alternatively, it may be desirable to give different shapes to the surfaces of the upper chamber region (330) and / or lower chamber region (332) of the chambers (320, 322, 324, 326). For example, it may be desirable to give the surfaces shapes that improve the manufacturability of each plate (300, 304), such as by enabling the molding of each plate (300, 304). In another embodiment, it may be desirable to give the surfaces of the chamber regions (330, 332) shapes that improve the pneumatic response to the region, such as by allowing the region to conform more closely to the surface in response to the pneumatic pressure (supplied, for example, via the pressure line (208)), while reducing or eliminating the strain on the region of the elastic layer (302) located within the chambers (320, 322, 324, 326) caused by the surface of the chamber region (330, 332), thereby allowing it to contact and seat more securely on the surface.

[0075] Figures 5 to 9B show an embodiment of a processing chip (400) that may provide the above-described features and functionalities. The processing chip (400) is the same as the processing chip (200) described above, unless otherwise stated below. In this regard, in combination with the rest of the system (100), the processing chip (400) may be used to provide in vitro synthesis, purification, concentration, preparation (e.g., encapsulation of therapeutic compositions into a delivery medium through a mixing process) and / or analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions. As shown in Figure 5, the processing chip (400) in this embodiment includes a plurality of fluid ports (420). Each fluid port (420) has an associated fluid channel (422) formed within the processing chip (400), and the fluid transmitted into the fluid port (420) flows through the corresponding fluid channel (422). As will be described in more detail below, each fluid port (420) is configured to receive fluid from the corresponding fluid line (206) from the fluid interface assembly (109). In this embodiment, each fluid channel (422) leads to a valve chamber (424), which is operable to selectively prevent or allow the fluid from the corresponding fluid channel (422) to be further transmitted along the processing chip (400), as will be described in more detail below.

[0076] As shown in Figure 5, the processing chip (400) in this embodiment includes several additional chambers (430, 450, 470) which can be used to serve various purposes during the process of producing therapeutic compositions as described herein. In just one example, such additional chambers (430, 450, 470) may be used to provide synthesis, purification, separation, compounding, and / or concentration of one or more therapeutic compositions, or to perform any other suitable function. Fluids may be transferred from one chamber (430) to another (430) via fluid connectors (432). In some versions, the fluid connectors (432) can operate like a valve between open and closed states (similar to, for example, a valve chamber (424)). In some other versions, the fluid connectors (432) remain open throughout the entire process of making the therapeutic compositions. In this embodiment, the chambers (430) are used to bring about the synthesis of polynucleotides, but the chambers (430) could instead serve any other suitable purpose.

[0077] In the embodiment shown in Figure 5, another valve chamber (434) is positioned between one of the chambers (430) and one of the chambers (450), thereby allowing the fluid to be selectively transferred from chamber (430) to chamber (450). The chambers (450) are provided in pairs and connected to one another, thereby allowing the processing tip (400) to transfer the fluid back and forth between the chambers (450). In this embodiment, a pair of chambers (450) are provided, but any other suitable number of chambers (450), including just one chamber (450) or three or more chambers (450), may be used. The chambers (450) may be used to provide fluid purification and / or serve any of the various other purposes described herein and may have any suitable configuration.

[0078] In versions where the chamber (450) is used for purification, the chamber (450) may include a material configured to absorb a selected portion from a fluid mixture within the chamber (450). In some such versions, this material may include a cellulose material that can selectively absorb double-stranded mRNA from the mixture. In some such versions, the cellulose material is inserted into only one of a pair of chambers (450), so that when the fluid from the first chamber (450) of this pair is mixed into the second chamber (450), mRNA and / or any other components may be effectively removed from the fluid mixture, which can then be transferred to another pair of chambers (470) further downstream for further processing or discharge. Alternatively, the chamber (450) may also be used for any other suitable purpose.

[0079] At least one additional valve chamber (452) is positioned between at least one chamber (450) and a corresponding chamber (470), thereby allowing fluid to be selectively transferred from chamber (450) to chamber (470) via the valve chamber (452). The chambers (470) are also connected to one another, and the processing tip (400) can transfer fluid back and forth between the chambers (470). The chambers (470) may be used to provide a mixture of fluids and / or may serve any of the various other purposes described herein and may have any suitable configuration.

[0080] As shown in Figure 5, the chamber (470) is also connected to an additional fluid port (421) via a corresponding fluid channel (423) and valve chamber (425). The fluid port (421), fluid channel (423), and valve chamber (425) can be configured and made operational in the same way as the aforementioned fluid port (420), fluid channel (422), and valve chamber (424). In some versions, the fluid port (421) is used to deliver additional fluid to the chamber (470). Furthermore, or instead, the fluid port (421) may be used to deliver fluid from the processing chip (400) to another device. For example, fluid from the chamber (470) may be delivered directly via the fluid port (421) to another processing chip (400), to one or more vials in a reagent storage frame (107), or to any other location.

[0081] The processing chip (400) further includes a plurality of reservoir chambers (460). In this embodiment, each reservoir chamber (460) is configured to receive and store fluid being transferred to or from the corresponding chambers (450, 470). Each reservoir chamber (460) has a corresponding inlet valve chamber (462) and an outlet valve chamber (464). Each inlet valve chamber (462) is positioned between the reservoir chamber (460) and the corresponding chambers (450, 470) and is operable to allow or prevent fluid flow between the reservoir chamber (460) and the corresponding chambers (450, 470). Each outlet valve chamber (464) is operable to meter the fluid flow between the reservoir chamber (460) and the corresponding fluid port (466). In some versions, each fluid port (466) is configured to transfer fluid from the corresponding vial in the reagent storage frame (107) to the corresponding reservoir chamber (460). Alternatively, each fluid port (466) may be configured to transfer fluid from the corresponding reservoir chamber (460) to the corresponding vial in the reagent storage frame (107). In this embodiment, the reservoir chamber (460) is used to provide metering of the fluid transferred to and / or from the processing tip (400). Alternatively, the reservoir chamber (460) may be used for any other suitable purpose, including but not limited to pressurizing the fluid transferred to and / or from the processing tip (400).

[0082] As shown in Figure 5, the processing chip (400) in this embodiment includes a plurality of pressure ports (440). Each pressure port (440) has an associated pressure channel (444) formed within the processing chip (400), and positively pressurized gas and / or negatively pressurized gas (e.g., vacuum) transmitted through the pressure port (440) is further transmitted through the corresponding pressure channel (444). As will be described in more detail below, each pressure port (440) is configured to receive pressurized gas from a corresponding pressure line (408) from the fluid interface assembly (109). In this embodiment, each pressure channel (444) leads to a corresponding chamber (424, 425, 430, 434, 450, 452, 460, 462, 464, 470), thereby providing valve action or peristaltic pump action through the chamber (424, 425, 430, 434, 450, 452, 460, 462, 464, 470), as will be described in more detail below.

[0083] In this embodiment, all fluid ports (421) and pressure ports (440) are located in the upper region of the processing chip (400), at least within the framework of Figure 6. In some other versions, all fluid ports (421) are located in the upper region of the processing chip (400), and all pressure ports (440) are located in the lower region of the processing chip (400). In yet another version, all fluid ports (421) are located in the lower region of the processing chip (400), and all pressure ports (440) are located in the upper region of the processing chip (400). In yet another variation, some fluid ports (421) may be located in the upper region of the processing chip (400), and others may be located in the lower region of the processing chip (400). Similarly, some pressure ports (440) may be located in the upper region of the processing chip (400), and other pressure ports (440) may be located in the lower region of the processing chip (400). Furthermore, or alternatively, some pressure ports (440) may be located directly above or below the corresponding chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470) (for example, along one or more ends of the processing chip (400) and not spaced apart from the corresponding chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470). In some versions, the pressure ports (440) arranged in this manner may be directly fluidically connected to the corresponding chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470) (for example, not fluidly connected to the corresponding chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470) via their respective pressure channels (444)).

[0084] The processing chip (400) may also 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 this regard, the processing chip (400) may include an electrically active region (not shown) similar to the electrically active region (212) that includes such electrical communication features.

[0085] As shown in Figure 6, the processing chip (400) further includes a first plate (500), an elastic layer (502), a second plate (504), and a third plate (506). As will be described in more detail below, some versions of the elastic layer (502) are in the form of a flexible film. The first plate (500) has an upper surface (410) and a lower surface (510), the lower surface (510) is placed alongside the elastic layer (502). The second plate (504) has an upper surface (512) and a lower surface (514), the upper surface (512) is placed alongside the elastic layer (502), and the lower surface (514) is placed alongside the third plate (506). Thus, the elastic layer (502) is placed between the first plate (500) and the second plate (504). Another elastic layer (not shown) similar to the elastic layer (316) may be placed between the second plate (504) and the third plate (506).

[0086] The plates (500, 504, 506) in this embodiment are substantially translucent to visible light and / or ultraviolet light. "Substantially translucent" means that at least 90% (and possibly 100%) of light is transmitted through the material compared to a translucent material. In some variations, one or more of the plates (500, 504, 506) may include a material that is substantially transparent to visible light and / or ultraviolet light. "Substantially transparent" means that at least 90% (and possibly 100%) of light is transmitted through the material compared to a completely transparent material. In another embodiment, one or more of the plates (500, 504, 506) may transmit ultraviolet light at a wavelength of about 260 nm with a transmittance ranging from about 0.2% to about 20%, including about 0.4% to about 15%, or about 0.5% to about 10%.

[0087] Furthermore, the plates (500, 504, 506) in this embodiment are rigid. In some other versions, one or more of the plates (500, 504, 506) are semi-rigid. The plates (500, 504, 506) may include glass, plastic, silicone, and / or any other suitable material. In some versions, one or more of the plates (500, 504, 506) are formed as a laminate of two or more layers of material, so that each plate (500, 504, 506) does not necessarily have to be formed as a single homogeneous continuum of material. The material constituting one of the plates (500, 504, 506) may be different from the material constituting the other plates (500, 504, 506).

[0088] Continuing to refer to Figure 6, the elastic layer (502) in this embodiment includes first and second liquid-impermeable flexible membranes (502a, 502b) stacked on top of each other in a vertically stacked arrangement. In the illustrated embodiment, the first flexible membrane (502a) is positioned below the second flexible membrane (502b), with the upper surface of the first flexible membrane (502a) juxtaposed with the lower surface of the second flexible membrane (502b). However, it will be understood that the first flexible membrane (502a) may instead be positioned above the second flexible membrane (502b), with the lower surface of the first flexible membrane (502a) juxtaposed with the upper surface of the second flexible membrane (502b). In some versions, one or both of the flexible membranes (502a, 502b) of the elastic layer (502) are gas-permeable despite being liquid-impermeable. In some such versions, certain regions of one or both of the flexible films (502a, 502b) of the elastic layer (502) are treated to be gas permeable, while the untreated regions of one or both of the flexible films (502a, 502b) of the elastic layer (502) are gas impermeable.

[0089] As will be described later, the elastic layer (502) may be used to drive fluid across the processing tip (400) via peristaltic pump action. Also as will be described later, the elastic layer (502) may be used to provide valves at various locations along the processing tip (400). In some versions, a single sheet of elastic material spans the entire width of the processing tip (200) to form the first flexible membrane (502a) of the elastic layer (302), and another single sheet of elastic material spans the entire width of the processing tip (200) to form the second flexible membrane (502b) of the elastic layer (302). In some other versions, two or more separate pieces of elastic material are used to form one or both of the flexible membranes (502a, 502b) of the elastic layer (302), and such separate pieces of elastic material are positioned at different locations across the width of the processing tip (200). In other versions, the first and second flexible films (502a, 502b) may be formed integrally with one another as a single component. For example, a single sheet of elastic material may span the entire width of the processed chip (200) and have a varying thickness to form a single, integral functional equivalent of both the first and second flexible films (502a, 502b) of the elastic layer (302). The varying thickness along this single, integral component can be provided by any suitable technique, including but not limited to molding, machining, and additive manufacturing. As just one example, one or both of the flexible films (502a, 502b) of the elastic layer (503) may include a polydimethylsilicone (PDMS) elastomer film.

[0090] In some versions, each flexible membrane (502a, 502b) may have a thickness of approximately 100 μm, and the elastic layer (502) may have an overall thickness of approximately 200 μm. It will be understood that the flexible membranes (502a, 502b) may have any other suitable thickness and may have different thicknesses from one another. The first flexible membrane (502a) is shown positioned below the second flexible membrane (502b), and the first flexible membrane (502a) may be called the lower flexible membrane (502a), and the second flexible membrane (502b) may be called the upper flexible membrane (502b), and the first flexible membrane (502a) may instead be positioned above the second flexible membrane (502b).

[0091] As best seen in Figures 9A-9C, the first plate (500) and the second plate (504) work together to define multiple chambers (520) (one shown), and the first flexible membrane (502a) of the elastic layer (502) bisects each chamber (520) into a corresponding upper chamber region (530) and a lower chamber region (532). As will be described in more detail below, during the operation of the processing tip (400), the upper chamber region (530) receives pressurized gas and the lower chamber region (532) receives fluid. Thus, the upper chamber region (530) can be considered a “dry” chamber region, and the lower chamber region (532) can be considered a “wet” chamber region. However, these “wet” and “dry” roles may be reversed in some modifications. In other words, some modifications of the processing chip (400) allow pressurized gas to be supplied to the lower chamber region (532), thereby constituting a “dry” chamber region, while the upper chamber region (530) receives fluid, thereby constituting a “wet” chamber region.

[0092] Any one or more of the chambers shown in Figure 5 (424, 425, 430, 434, 450, 452, 460, 462, 464, 470) can be configured and operated in the same way as the chamber (520) shown in Figures 9A to 9C. For example, chamber (520) may be similar to one or more of the chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470). In some versions, chamber (520) may be similar to one or more of the valve chambers (424, 425, 434, 452, 462, 464). In this regard, the chamber (520) may have a generally circular aerial profile (e.g., the profile of the chamber (520) when viewed from above) and may extend perpendicularly along a central longitudinal axis (L) oriented generally perpendicular to the plates (500, 502). In some versions, the chamber (520) may be configured to provide ventilation, such as a vacuum cap as described above. Thus, it should be understood that the chamber (520) may be configured to perform any of the functions of the fluid receiving areas of the processing tips (200, 400) described herein.

[0093] The fluid ports (420) may be formed through the first plate (500) shown in Figure 6. As shown in Figures 7 and 8, the corresponding openings (542a, 542b) are formed through the regions of the flexible membrane (502a, 502b) of the elastic layer (502) beneath each fluid port (420). One of the fluid channels (422) extends from each opening (542a, 542b) to the lower chamber region (532) of the chamber (520). As previously stated, each fluid port (420) is configured to receive a fluid line (206) from the fluid interface assembly (109). The distal end of the fluid line (206) may be configured to seal in contact with the region of the elastic layer (502) exposed by the fluid port (420) and to transmit fluid through each opening (542a, 542b). In some versions, a spring or other elastic member provides elastic bias to the fluid line (206), pressing the distal end of the fluid line (206) against the region of the elastic layer (502) exposed by the fluid port (420), thereby maintaining a seal. Fluid from the fluid line (206) reaches the lower chamber region (532) of the chamber (520) via the fluid channel (422). As will be described in more detail below, this fluid can be further transferred from the chamber (520) to other chambers or other features within the processing tip (400), to storage vials within the reagent storage frame (107), or otherwise processed through peristaltic pump action provided via the elastic layer (502). It should also be understood that any of the other fluid ports (421, 466) shown in Figure 5 can be configured and made operational in a similar manner to the fluid port (420). As shown in Figures 7 and 8, the corresponding openings (543a, 543b) are formed through the regions of the flexible membrane (502a, 502b) of the elastic layer (502) located beneath the respective fluid ports (421, 466).

[0094] The pressure ports (440) may be formed through the first plate (500) shown in Figure 6. As shown in Figures 7 and 8, the corresponding openings (544a, 544b) are formed through the regions of the flexible membrane (502a, 502b) of the elastic layer (502) beneath each pressure port (440). One of the pressure channels (444) extends from each opening (544a, 544b) to the upper chamber region (530) of the first chamber (520). As previously stated, each pressure port (440) is configured to receive a pressure line (208) from the fluid interface assembly (109) and thereby receive pressurized gas from the pressure source (117). The distal end of the pressure line (208) is configured to seal in contact with a region of the elastic layer (502) exposed by the pressure port (440) and to transmit positively pressurized or negatively pressurized gas through the respective openings (544, 544b). In some versions, a spring or other elastic member provides elastic bias to the pressure line (208), pressing the distal end of the pressure line (208) against the region of the elastic layer (502) exposed by the pressure port (440), thereby maintaining the seal. The positively pressurized or negatively pressurized gas from the pressure line (208) reaches the upper chamber region (530) of the chamber (520) via the pressure channel (444).

[0095] Only one pressure line (208) may be connected to the processing chip (400), or the processing chip (400) may have multiple connected pressure lines (208), such pressure lines (208) will be recognized as independently applying positive or negative pressure to the corresponding chambers (520) of the processing chip (400). In some versions, one or more of the chambers (520) have their own dedicated pressure lines (208) and corresponding pressure channels (444). Furthermore, or alternatively, one or more of the chambers (520) may share a common pressure line (208) via the same pressure channel (444) or via separate pressure channels (444). In some versions, the pressure channels (444) may be formed through a second plate (504). Furthermore, or alternatively, some pressure channels (444) (or regions of pressure channels (444)) may be formed by a first plate (500). For example, several pressure channels (444) (or regions of pressure channels (444)) may be formed between a recess on the lower surface of the first plate (500) and the upper surface of the elastic layer (502).

[0096] As described above, the first flexible membrane (502a) of the elastic layer (502) extends across each chamber (520) at or near its vertical midpoint, thereby bisecting each chamber (520) and fluidly separating the corresponding upper chamber region (530) and lower chamber region (532) from each other, at least when the elastic layer (502) is not deflected. In this regard, the first flexible membrane (502a) of the elastic layer (502) can be configured and made operable in the same way as the elastic layer (302) described above, but the first flexible membrane (502a) may have a thinner thickness than the elastic layer (302) in some versions. For example, the first flexible membrane (502a) of the elastic layer (502) may be configured to deflect within each chamber (520) as described above in relation to Figures 4A to 4F, thereby driving one or more fluids through each chamber (520).

[0097] Conversely, the second flexible membrane (502b) of the elastic layer (502) is not configured to bisect at least one chamber (520), nor to extend across at least one chamber (520), nor to deflect within at least one chamber (520). In some versions, the second flexible membrane (502b) of the elastic layer (502) does not need to bisect any of the chambers (520), nor to extend across any of the chambers (520), nor to deflect within any of the chambers (520). In this regard, the second flexible membrane (502b) of the elastic layer (502) may be substantially identical to the first flexible membrane (502a) (for example, when viewed from above), except that the second flexible membrane (502b) further includes a plurality of openings (e.g., notches) (540), each of which is configured to be perpendicular to the respective chamber (520), and each opening (540) has a roughly defined center point with respect to its respective longitudinal axis (L). For example, each opening (540) may be located directly below the corresponding upper chamber region (530) and / or directly above the corresponding lower chamber region (532). Figure 8 best illustrates the notches in the second flexible membrane (502b) of the elastic layer (502).

[0098] Each opening (540) may be sized and molded similarly to its respective chamber (520) (for example, when viewed from above), and the periphery of each opening (540) may generally follow the periphery of its respective chamber (520). For example, each opening (540) corresponding to a chamber (520) having a generally circular aerial profile (for example, the corresponding chamber (520) is similar to one or more of the valve chambers (424, 425, 434, 452, 462, 464) may also have a generally circular aerial profile; each opening (540) corresponding to a chamber (520) having a generally oval aerial profile (for example, the corresponding chamber (520) is similar to one or more of the reservoir chambers (460)) may also have a generally oval aerial profile; Each opening (540) corresponding to a chamber (520) having a generally rectangular aerial profile with rounded corners (for example, the corresponding chamber (520) is similar to one or more of the chambers (430, 470)) may also have a generally rectangular aerial profile with rounded corners; each opening (540) corresponding to a chamber (520) having a generally rectangular aerial profile with concave sides (for example, the corresponding chamber (520) is similar to one or more of the chambers (450)) may also have a generally rectangular aerial profile with concave sides. Thus, each opening (540) of the second flexible membrane (502b) may be directly above or directly below the corresponding portion of the first flexible membrane (502a) configured to deflect within each chamber (520). The opening (540) can accommodate the deflection of the first flexible membrane (502a) by preventing the second flexible membrane (502b) of the elastic layer (502) from forming wrinkles or interfering in a different way with the deflection of the first flexible membrane (502a) of the elastic layer (502) within the chamber (520).

[0099] In some versions, one or more openings (540) may partially extend into each chamber (520). For example, the periphery of each opening (540) may be positioned at least about 50 μm inward from the periphery of each chamber (520). In another embodiment, the periphery of each opening (540) may be positioned at least about 200 μm inward from the periphery of each chamber (520). Such spacing may help ensure proper adhesion between the elastic layer (502) and the plate (500) in the region surrounding each chamber (520).

[0100] From the above, it should be understood that the combination of flexible membranes (502a, 502b) can increase the thickness of the elastic layer (502) in regions where the elastic layer (502) maintains a constant interface with the plate (500), thereby meeting the requirement to strengthen adhesion at these interfaces. Furthermore, the presence of the opening (540) and flexible membrane (502a) in the chamber (520) region allows for a reduction in the thickness of the elastic layer (502) in the chamber (520) region, thereby meeting the requirement to enhance the effectiveness of valve action and peristaltic pump action within the chamber (520) (which, as described above, can be configured as one or more of any other type of fluid receiving regions within a valve chamber, synthesis chamber, purification chamber, mixing chamber, metering chamber, vacuum cap, or processing tip (200, 400)). In other words, the elastic layer (502) can simultaneously meet two requirements that would otherwise seem contradictory.

[0101] As shown in Figures 9A to 9C, the upper chamber region (530) and the lower chamber region (532) of the chamber (520) are each contoured to exhibit a generally flat, bell-shaped cross-sectional profile. More specifically, the upper chamber region (530) is defined by a generally annular radial outer surface (530a), a generally annular radial middle surface (530b), and a generally circular radial inner surface (530c). The outer surface (530a) and the middle surface (530b) are curved, while the inner surface (530c) is flat. In this regard, the outer surface (530a) extends convexly from the radial outer circumference of the chamber (520) radially inward and upward on the first side of the longitudinal axis (L); the intermediate surface (530b) extends concavely from the radially inward and upward from the radially inward end of the outer surface (530a), thereby the outer surface (530a) and the intermediate surface (530b) together exhibit a roughly S-shaped cross-sectional profile on the first side of the longitudinal axis (L) (for example, within the framework of Figures 9A to 9C); and the inner surface (530c) extends across the longitudinal axis (L) from the radially inward end of the intermediate surface (530b) to the surfaces (410, 510, 512, 514) of the plates (500, 504). The inner surface (530c) extends generally parallel to the and / or the flexible film (502a, 502b) of the elastic layer (502), thereby determining the approximate center point of the inner surface (530c) with respect to the longitudinal axis (L); the intermediate surface (530b) extends concavely radially outward and downward from the end of the inner surface (530c) on the second side of the longitudinal axis (L); and the outer surface (530a) extends convexly radially outward and downward from the radially outer end of the intermediate surface (530b), thereby the outer surface (530a) and the intermediate surface (530b) collectively exhibit a roughly inverted S-shaped cross-sectional profile on the second side of the longitudinal axis (L) (for example, within the framework of Figures 9A to 9C).

[0102] As described above, the inner surface (530c) is circular and has a roughly defined center point with respect to the longitudinal axis (L). The outer surface (530a) and the intermediate surface (530b) may also each have roughly defined center points with respect to the longitudinal axis (L), and each may have a uniform configuration in the circumferential direction with respect to the longitudinal axis (L), thereby allowing the upper chamber region (530) to be roughly symmetrical with respect to the longitudinal axis (L). For example, the outer surface (530a) may have a constant radius of curvature and / or a constant arc length, and the intermediate surface (530b) may also have a constant radius of curvature and / or a constant arc length. In some versions, the outer surface (530a) may have a first radius of curvature and / or a first arc length, and the intermediate surface (530b) may have a second radius of curvature different from the first radius of curvature and / or a second arc length different from the first arc length.

[0103] Similarly, the lower chamber region (532) is defined by a generally annular radial outer surface (532a), a generally annular radial middle surface (532b), and a generally circular radial inner surface (532c). The outer surface (532a) and the middle surface (532b) are curved, respectively, while the inner surface (532c) is flat. In this regard, the outer surface (532a) extends convexly from the radial outer circumference of the chamber (520) radially inward and downward on the first side of the longitudinal axis (L); the intermediate surface (532b) extends concavely from the radially inward and downward from the radially inward end of the outer surface (532a), thereby the outer surface (532a) and the intermediate surface (532b) together exhibit a roughly inverted S-shaped cross-sectional profile on the first side of the longitudinal axis (L) (for example, within the framework of Figures 9A to 9C); and the inner surface (532c) extends across the longitudinal axis (L) from the radially inward end of the intermediate surface (532b) to the surfaces (410, 510, 512, 51) of the plates (500, 504). 4) extends generally parallel to and / or generally parallel to the flexible film (502a, 502b) of the elastic layer (502), thereby determining the approximate center point of the inner surface (532c) with respect to the longitudinal axis (L); the intermediate surface (532b) extends concavely radially outward and upward from the end of the inner surface (532c) on the second side of the longitudinal axis (L); and the outer surface (532a) extends convexly radially outward and upward from the radially outer end of the intermediate surface (532b), thereby the outer surface (532a) and the intermediate surface (532b) collectively exhibit a roughly S-shaped cross-sectional profile on the second side of the longitudinal axis (L) (for example, within the framework of Figures 9A to 9C).

[0104] As described above, the inner surface (532c) is circular and has a roughly defined center point with respect to the longitudinal axis (L). The outer surface (532a) and the intermediate surface (532b) may also each have roughly defined center points with respect to the longitudinal axis (L), and each may have a uniform configuration in the circumferential direction with respect to the longitudinal axis (L), thereby allowing the lower chamber region (532) to be roughly symmetrical with respect to the longitudinal axis (L). For example, the outer surface (532a) may have a constant radius of curvature and / or a constant arc length, and the intermediate surface (532b) may also have a constant radius of curvature and / or a constant arc length. In some versions, the outer surface (532a) may have a first radius of curvature and / or a first arc length, and the intermediate surface (532b) may have a second radius of curvature different from the first radius of curvature and / or a second arc length different from the first arc length.

[0105] Furthermore, or alternatively, the chamber (520) may be substantially symmetric with respect to the elastic layer (502). For example, the outer surface (532a) of the lower chamber region (532) may have the same radius of curvature, arc length, outer diameter, and / or inner diameter as the outer surface (530a) of the upper chamber region (530); the intermediate surface (532b) of the lower chamber region (532) may have the same radius of curvature, arc length, outer diameter, and / or inner diameter as the intermediate surface (530b) of the upper chamber region (530); and / or the inner surface (532c) of the lower chamber region (532) may have the same diameter as the inner surface (530c) of the upper chamber region (530). In some other versions, the chamber (520) may be substantially asymmetric with respect to the elastic layer (502). For example, one of the upper chamber region (530) or the lower chamber region (532), for example the lower chamber region (532), may have the illustrated flat bell-shaped cross-sectional profile, while the other of the upper chamber region (530) or the lower chamber region (532), for example the upper chamber region (530), may have a completely different cross-sectional profile, such as an arc-shaped cross-sectional profile, a semicircular cross-sectional profile, a rectangular cross-sectional profile, and so on.

[0106] As described above, the elastic layer (502) can operate to drive the fluid through the processing tip (400) by peristaltic pump action and to stop the movement of the fluid through the processing tip (400) by providing valve action. An example of such operation is illustrated in the sequence drawn in Figures 9A to 9C. In this embodiment, the chamber (520) functions as a valve chamber (for example, the chamber (520) is analogous to one or more of the valve chambers (424, 425, 434, 452, 462, 464)). This configuration, arrangement, and use of the chamber (520) is provided as an exemplary embodiment. The chamber (520) may instead be configured, arranged, and used in other ways (for example, the chamber (520) is analogous to the reservoir chamber (460) and / or one or more of the chambers (430, 450, 470)).

[0107] Figure 9A shows the processing tip (400) before the fluid has been transferred to it and before the pressurized gas has been transferred to it. In Figure 9B, the positively pressurized gas is transferred to the upper chamber region (530) of the chamber (520). In this state, the positively pressurized gas deforms the portion of the first flexible membrane (502a) of the elastic layer (502) inside the chamber (520), causing the first flexible membrane (502a) of the elastic layer (502) to be in contact with and fully seated against the surface (532a, 532b, 532c) of the lower chamber region (532) of the chamber (520). The first flexible membrane (502a) of the elastic layer (502) is in contact with the surfaces (532a, 532b, 532c) of the lower chamber region (532) of the chamber (520) and is seated in this manner, preventing fluid from entering the chamber (520), so that the chamber (520) acts like a shut-off valve in the state shown in Figure 9B.

[0108] In Figure 9C, the negatively pressurized gas is transferred to the upper chamber region (530) of the chamber (520). In this state, the negatively pressurized gas in the upper chamber region (530) of the chamber (520) deforms the corresponding portion of the first flexible membrane (502a) of the elastic layer (502) inside the chamber (520), causing it to contact and fully seat against the surface (530a, 530b, 530c) of the upper chamber region (530) of the chamber (520). This effectively opens the valve formed by the chamber (520), allowing the chamber (520) to receive fluid (not shown) and occupy the entire volume of the chamber (520). Subsequently, positively pressurized gas is transferred to the upper chamber region (530) of the chamber (520), deforming the first flexible membrane (502a) back to the state shown in Figure 9B, thereby allowing the fluid to be released from the chamber (520).

[0109] As described above, the first flexible membrane (502a) of the elastic layer (502) may have a thinner thickness than the aforementioned elastic layer (302). Since there is an opening (540) in the region of the elastic layer (502) located within the chamber (520), the elastic layer (502) is thinner in that region than the elastic layer (302), thereby improving the response of that region of the elastic layer (502) to pneumatic pressure. For example, the elastic layer (502) may provide improved valve action and / or pump action, at least compared to the elastic layer (302). In addition, or instead, the valve action and / or pump action provided by the elastic layer (502) may be achieved by reduced pneumatic pressure, at least compared to the pneumatic pressure required to operate the elastic layer (302) pneumatically. Such reduced pneumatic pressure may be relatively easy to maintain and / or result in improved safety. The ability of the elastic layer (502) to be pneumatically actuated at reduced air pressure also allows one or more of the chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470) to have a reduced size compared to at least the chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270), thereby saving space on the processing chip (400). In some versions, the valve and / or pump action provided by the elastic layer (502) can be achieved via an air pressure of approximately 34.47–41.37 kPa (approximately 5–6 psi). In another embodiment, the valve and / or pump action provided by the elastic layer (502) can be achieved via an air pressure of approximately 27.58 kPa (approximately 4 psi).

[0110] Furthermore, or alternatively, the first flexible film (502a) and the second flexible film (502b) may, collectively, have an overall thickness greater than that of the elastic layer (302) described above. Since there is no opening (540) in the region of the elastic layer (502) bonded to the first plate (500) and / or the second plate (504), the elastic layer (502) is thicker than the elastic layer (302) in that region, thereby improving the seal between that region of the elastic layer (502) and one or both of the first plate (500) and / or the second plate (504). For example, the upper and lower surfaces of the elastic layer (502) may exhibit improved conformity to the respective juxtaposed surfaces (510, 512) of the plates (500, 504) at least compared to the elastic layer (302). In addition, or instead, the upper and lower surfaces of the elastic layer (502) may exhibit reduced deformability to any of the channels (422, 423, 444), chambers (424, 425, 430, 434, 450, 452, 460, 462, 464, 470), and / or other microfluidic path features of the processing chip (400) during assembly, at least compared to the elastic layer (302). In this regard, the ratio of the thickness of the elastic layer (502) to the width of any of the channels (422, 423, 444) can be approximately 1:2 or greater. In other words, the relationship between the thickness of the elastic layer (502) and the width of any of the channels (422, 423, 444) can be expressed as follows:

number

[0111] It will be recognized that the flat bell shape of the upper chamber region (530) and lower chamber region (532) of the chamber (520) can improve the manufacturability of each plate (500, 504) by enabling the molding of each plate (500, 504). In addition, or instead, the shapes of the upper chamber region (530) and the lower chamber region (532) of the chamber (520) can result in improved pneumatic response in those regions, at least compared to the surfaces of the chamber regions (330, 332), by allowing the regions of the elastic layer (502) placed within the chamber (520) to conform more closely to the respective surfaces (530a, 530b, 530c, 532a, 532b, 532c) of the chamber regions (530, 532) in response to the pneumatic pressure (supplied, for example, via the pressure line (208)), thereby enabling them to contact and seat more completely with the respective surfaces (530a, 530b, 530c, 532a, 532b, 532c).

[0112] In some cases, each surface (530a, 530b, 530c, 532a, 532b, 532c) can prevent the corresponding region of the elastic layer (502) from being deformed when it sits in contact with them. In this regard, the surfaces (530a, 530b, 530c, 532a, 532b, 532c) can be sized and constructed based on a predetermined deformation shape of the corresponding region of the elastic layer (502) in response to a predetermined air pressure. For example, the deformation shape of a particular region of the elastic layer (502) can be determined by simulating the deformation of the region of the elastic layer (502) in response to a predetermined air pressure. The surfaces (530a, 530b, 530c) of the upper chamber region (530) and / or the surfaces (532a, 532b, 532c) of the lower chamber region (532) may be sized and configured to collectively fit (e.g., complement) the deformation shape of the corresponding regions of the elastic layer (502). This allows the corresponding regions of the elastic layer (502) to contact and precisely seat on the surfaces (530a, 530b, 530c, 532a, 532b, 532c) in response to air pressure, while reducing or eliminating the strain on those regions of the elastic layer (502) caused by the surfaces (530a, 530b, 530c, 532a, 532b, 532c). As a result, the surfaces (530a, 530b, 530c, 532a, 532b, 532c) can provide an improved seal with the elastic layer (502) and / or achieve a sufficient seal with the elastic layer (502) at a reduced air pressure.

[0113] Figure 10 shows an example of method (600) for designing a chamber (520). Although method (600) is described in the context of designing a chamber (520) used for valve operation, it will be understood that method (600) can also be used for designing a chamber (520) used for pump operation.

[0114] Method (600) begins with step (601), in which the material and thickness of an elastic layer, such as elastic layer (302) or elastic layer (502), are selected. For example, the material may be selected as PDMS and / or the thickness may be selected as approximately 200 μm. If elastic layer (502) is used, the material and / or thickness of elastic layer (502) selected in step (601) may be the material and / or thickness of the first flexible film (502a) of elastic layer (502). Method (600) proceeds from step (601) to step (602), in which the diameter of a chamber, such as chamber (520), is selected. For example, the diameter may be selected as approximately 2.1 mm. Method (600) proceeds from step (602) to step (603), in which the depth of an adjacent channel, such as channel (422), is selected. For example, the depth may be selected as approximately 200 μm. Method (600) proceeds from step (603) to step (604), in which a target pressure for activating the elastic layer (for example, to close the chamber) is selected. For example, the target pressure may be selected as approximately 27.58 kPa (approximately 4 psi).

[0115] Method (600) proceeds from step (604) to step (605), in which the application of increased pressure to the elastic layer is simulated. For example, it may be simulated that the pressure applied to the elastic layer increases from approximately 0 kPa (approximately 0 psi) to approximately 103.42 kPa (approximately 15 psi). Method (600) proceeds from step (605) to step (606), in which it is determined whether the deflected depth of the elastic layer simulated at the target pressure is sufficient to perform the desired valve action (for example, by being able to close the adjacent channel). For example, the deflected depth of the elastic layer may be determined to be sufficient if it is above a predetermined threshold, such as when the deflected depth of the elastic layer is approximately twice the depth of the adjacent channel (e.g., approximately 400 μm). If it is determined that the deflected depth of the elastic layer is insufficient, the method returns from step (606) to step (602), in which a larger chamber diameter may be selected. If the deflected depth of the elastic layer is determined to be sufficient, the method proceeds from step (606) to step (607), in which the deformation shape of the elastic layer simulated at the target pressure is applied to generate the shape of the chamber, for example, the shape of the upper chamber region (530) and / or lower chamber region (532) of chamber (520). For example, the specific size and configuration of the surfaces (530a, 530b, 530c, 532a, 532b, 532c) may be generated based on the deformation shape of the elastic layer simulated at the target pressure.

[0116] The specific size and configuration of the surfaces (530a, 530b, 530c, 532a, 532b, 532c) based on a given air pressure can be scaled (e.g., enlarged or reduced) according to various other parameters of the processing chip (400), such as the depth of any of the channels (422, 423, 444) and / or the thickness of the first flexible film (502a) of the elastic layer (502). Thus, the flat bell shape of the upper chamber region (530) and / or lower chamber region (532) can be fine-tuned to suit various applications.

[0117] Figure 11 shows another embodiment of a processing chip (700) which is similar to the processing chip (400) described above unless otherwise specified. As shown in the figure, the processing chip (700) includes a first plate (800), an elastic layer (802), and a second plate (804). The first plate (800) has an upper surface (710) and a lower surface (810), the lower surface (810) being placed alongside the elastic layer (802). The second plate (804) has an upper surface (812) and a lower surface (814), the upper surface (812) being placed alongside the elastic layer (802). Thus, the elastic layer (802) is placed between the first plate (800) and the second plate (804). The elastic layer (802) of this embodiment includes a first liquid-impermeable flexible membrane (802a) and a second liquid-impermeable flexible membrane (802b) stacked on top of each other in a vertically stacked arrangement. The first flexible membrane (802a) may be called the lower flexible membrane (802a), and the second flexible membrane (802b) may be called the upper flexible membrane (802b). The first plate (800) and the second plate (804) cooperate to define a plurality of chambers (720), and the second flexible membrane (802b) of the elastic layer (802) bisects each chamber (820) into a corresponding upper chamber region (830) and a lower chamber region (832). Fluid can be transmitted between at least one lower chamber region (832) and one or more fluid ports (not shown) via associated fluid channels (722), and can also be transmitted from one lower chamber region (832) to another lower chamber region (832) via fluid channels (732).

[0118] In the illustrated embodiment, the first flexible membrane (802a) of the elastic layer (802) is substantially identical to the second flexible membrane (802b) (when viewed, for example, from above), except that the first flexible membrane (802a) further includes a plurality of openings (e.g., notches) (840) each configured to be aligned perpendicularly with each chamber (820). For example, each opening (840) may be located directly below the corresponding upper chamber region (830) and / or directly above the corresponding lower chamber region (832).

[0119] Therefore, unlike the embodiments shown in Figures 5 to 9C, in which the opening (540) is incorporated into the second flexible membrane (502b), Figure 11 shows the opening (840) incorporated into the first flexible membrane (502a). In some modifications, the opening (840) can be incorporated into both flexible membranes (802a, 802b) at various positions on the elastic layer (802) corresponding to various chambers (820). For example, the first flexible membrane (802a) may include an opening (840) aligned perpendicularly to the first chamber (820), while the second flexible membrane (802b) may not include an opening (840) aligned perpendicularly to the first chamber (820); the second flexible membrane (802b) may include an opening (840) aligned perpendicularly to the second chamber (820), while the first flexible membrane (802a) may not include an opening (840) aligned perpendicularly to the second chamber (820). In other words, the opening (840) can be repeatedly incorporated into the first flexible membrane (802a) and then into the second flexible membrane (802b).

[0120] VI. Examples of combinations The following embodiments relate to various non-inclusive methods to which the teachings herein may be combined or applied. It should be understood that the following embodiments are not intended to limit the scope of any claims that may be presented at any time in this application or a subsequent application thereof. This is not intended as a disclaimer. The following embodiments are provided solely for illustrative purposes. Various teachings herein are intended to be composed of and applied in many other ways. It is also intended that some modifications may omit certain features mentioned in the following embodiments. Therefore, no aspect or feature mentioned below should be considered important unless explicitly indicated at a later date as important by the inventor or a successor relating to the inventor. If any claim containing additional features other than those mentioned below is presented in this application or a subsequent application relating to this application, those additional features shall not be presumed to have been added for any reason relating to patentability.

[0121] Example 1 A microfluidic device comprising: (a) a first plate; (b) a second plate; (c) a microfluidic path defined between the first plate and the second plate, the microfluidic path comprising at least one chamber; and (d) an elastic layer disposed between the first plate and the second plate, the elastic layer comprising: (i) a first membrane extending across at least one chamber, the first membrane being configured to deflect to drive fluid through at least one chamber; and (ii) a second membrane having at least one opening, the at least one opening being aligned with at least one chamber.

[0122] Example 2 The microfluidic apparatus according to Example 1, wherein the second membrane does not extend across at least one chamber.

[0123] Example 3 A microfluidic apparatus according to any one of Examples 1 to 2, wherein at least one chamber extends along a central longitudinal axis, and at least one opening has a center point defined with respect to the central longitudinal axis.

[0124] Example 4 The microfluidic apparatus according to any one of Examples 1 to 3, wherein the first membrane is configured to fluidly isolate a first region of at least one chamber from a second region of at least one chamber.

[0125] Example 5 The microfluidic apparatus according to Embodiment 4, wherein at least one opening is located directly above the first region and directly below the second region.

[0126] Example 6 A microfluidic apparatus according to any one of Examples 1 to 5, wherein the first membrane and the second membrane are stacked on top of each other.

[0127] Example 7 A microfluidic apparatus according to any one of Examples 1 to 6, wherein one of the first or second membranes is bonded to a first plate, and the other of the first or second membranes is bonded to a second plate.

[0128] Example 8 A microfluidic apparatus according to any one of Examples 1 to 7, wherein at least one of the first or second membrane comprises polydimethylsilicone (PDMS).

[0129] Example 9 A microfluidic apparatus according to any one of Examples 1 to 8, wherein at least one of the first or second membrane has a thickness of about 100 μm.

[0130] Example 10 The microfluidic apparatus according to any one of Examples 1 to 9, wherein the elastic layer has an overall thickness of approximately 200 μm.

[0131] Example 11 A microfluidic apparatus according to any one of Examples 1 to 10, wherein at least one chamber and at least one opening have the same profile as each other.

[0132] Example 12 The microfluidic apparatus according to Example 11, wherein the profile is at least one of circular, oval, or at least partially rectangular.

[0133] Example 13 A microfluidic apparatus according to any one of Examples 1 to 12, wherein at least one chamber includes a valve chamber.

[0134] Example 14 A microfluidic apparatus according to any one of Examples 1 to 13, wherein at least one chamber comprises a plurality of chambers, and at least one opening comprises a plurality of openings, and each of the plurality of openings is aligned with a corresponding chamber of the plurality of chambers.

[0135] Example 15 A microfluidic apparatus according to Example 14, wherein multiple openings have different profiles from one another.

[0136] Example 16 A microfluidic device comprising: (a) an upper plate; (b) a lower plate; (c) a microfluidic path defined between the upper plate and the lower plate, the microfluidic path comprising a chamber; and (d) an elastic layer disposed between the upper plate and the lower plate, the elastic layer comprising: (i) a first membrane that bisects the chamber so as to define an upper chamber region and a lower chamber region, the first membrane being configured to deflect to drive fluid through the chamber; and (ii) a second membrane having an opening, the opening being located directly below the upper chamber region and directly above the lower chamber region.

[0137] Example 17 The microfluidic apparatus according to Example 16, wherein the first and second films are stacked on top of each other.

[0138] Example 18 The microfluidic apparatus according to any one of Examples 16 to 17, wherein the first membrane is located below the second membrane.

[0139] Example 19 A microfluidic apparatus according to any of Examples 16 to 18, wherein the chamber and opening have the same profile as each other.

[0140] Example 20 A microfluidic device comprising: (a) a first plate; (b) a second plate; (c) a microfluidic path defined between the first plate and the second plate, the microfluidic path comprising a chamber; and (d) an elastic layer disposed between the first plate and the second plate, the elastic layer comprising: (i) a first membrane configured to fluidly isolate a first region of the chamber from a second region of the chamber, the first membrane configured to deflect to drive fluid through the chamber; and (ii) a second membrane having an opening, the opening being aligned with the chamber.

[0141] Example 21 A microfluidic device comprising: (a) a first plate; (b) a second plate; (c) an elastic layer disposed between the first plate and the second plate; and (d) a microfluidic path defined between the first plate and the second plate, the microfluidic path comprising a chamber, the chamber extending along a central longitudinal axis, and at least a portion of the chamber having a flat, bell-shaped cross-sectional profile.

[0142] Example 22 The microfluidic apparatus according to Example 21, wherein the elastic layer extends across the chamber to define a first region and a second region of the chamber, and at least one of the first region or the second region has a flat, bell-shaped cross-sectional profile.

[0143] Example 23 The microfluidic apparatus according to Example 22, wherein each of the first and second regions has a flat, bell-shaped cross-sectional profile.

[0144] Example 24 The microfluidic apparatus according to any one of Examples 21 to 23, wherein the chamber is substantially symmetrical with respect to the central longitudinal axis.

[0145] Example 25 The microfluidic apparatus according to any one of Examples 21 to 24, wherein the chamber includes a radially inner surface, a radially outer surface, and a radially intermediate surface that collectively define a flat, bell-shaped cross-sectional profile.

[0146] Example 26 The microfluidic apparatus according to Example 25, wherein the radially inner surface has a flat cross-sectional profile.

[0147] Example 27 A microfluidic apparatus according to any one of Examples 25 to 26, wherein the radially outer surface and the radially intermediate surface each have a curved cross-sectional profile.

[0148] Example 28 The microfluidic apparatus according to Example 27, wherein the radially outer surface has a convex cross-sectional profile.

[0149] Example 29 The microfluidic apparatus according to any one of Examples 27 to 28, wherein the radial intermediate surface has a concave cross-sectional profile.

[0150] Example 30 The microfluidic apparatus according to any one of Examples 27 to 29, wherein the radially outer surface and the radially intermediate surface collectively define at least one of an S-shaped cross-sectional profile or an inverted S-shaped cross-sectional profile.

[0151] Example 31 The microfluidic apparatus according to any one of Examples 25 to 30, wherein the radially inner surface is generally circular.

[0152] Example 32 The microfluidic apparatus according to Example 31, wherein the radially outer surface and the radially intermediate surface are each generally annular.

[0153] Example 33 A microfluidic apparatus according to any one of Examples 25 to 32, wherein a portion of the elastic layer is configured to deflect to drive a fluid through the chamber, and this portion of the elastic layer is configured to conform to the radially inner surface, the radially outer surface, and the radially intermediate surface, respectively, when deflected.

[0154] Example 34 The microfluidic apparatus according to Example 33, comprising: (i) a first membrane extending across a chamber, the first membrane being configured to deflect to drive fluid through the chamber, and the first membrane being configured to conform to the radially inner surface, the radially outer surface, and the radially intermediate surface, respectively, when deflected; and (ii) a second membrane having an opening, the opening being aligned with the chamber.

[0155] Example 35 The chamber is a microfluidic device according to any one of Examples 21 to 34, including a valve chamber.

[0156] Example 36 A microfluidic apparatus comprising: (a) a first plate; (b) a second plate; (c) an elastic layer disposed between the first plate and the second plate; and (d) a microfluidic path defined between the first plate and the second plate, the microfluidic path comprising a chamber, the chamber extending along a central longitudinal axis, the chamber comprising: (i) a radially inner surface having a flat cross-sectional profile; (ii) a radially outer surface having a convex cross-sectional profile; and (iii) a radially intermediate surface having a concave cross-sectional profile.

[0157] Example 37 The microfluidic apparatus according to Example 36, wherein the radially outer surface and the radially intermediate surface collectively define at least one of an S-shaped cross-sectional profile or an inverted S-shaped cross-sectional profile.

[0158] Example 38 The microfluidic apparatus according to any one of Examples 36 to 37, wherein the radially inner surface is generally circular.

[0159] Example 39 The microfluidic apparatus according to Example 38, wherein the radially outer surface and the radially intermediate surface are each generally annular.

[0160] Example 40 A microfluidic apparatus comprising: (a) a first plate; (b) a second plate; (c) an elastic layer disposed between the first plate and the second plate; and (d) a microfluidic path defined between the first plate and the second plate, the microfluidic path comprising a chamber, the chamber extending along a central longitudinal axis, the chamber comprising: (i) a radially inner surface; (ii) a radially outer surface; and (iii) a radially intermediate surface, wherein (A) the radially outer surface extends convexly from the radially outer circumference to the radially inward side of the chamber on the first side of the longitudinal axis; and (B) the radially intermediate surface is A microfluidic device comprising: (C) a microfluidic path extending concavely inward from the radially inner end of the radially outer surface on the first side of the longitudinal axis; (D) a microfluidic path extending concavely outward from the end of the radially inner surface on the second side of the longitudinal axis across the longitudinal axis; and (E) a microfluidic path extending convexly outward from the radially outer end of the radially outer surface on the second side of the longitudinal axis.

[0161] VII. Others The foregoing description is provided to enable those skilled in the art to implement the various configurations described herein. While the subject art is described in detail with reference to various drawings and configurations, these are for illustrative purposes only and should not be interpreted as limiting the scope of the subject art.

[0162] There are many other ways to implement the subject art. Various functions and elements described herein may be divided in ways different from those illustrated without departing from the scope of the subject art. Various modifications to these embodiments may be readily apparent to those skilled in the art, and the general principles defined herein may also apply to other embodiments. Thus, many changes and modifications can be made to the subject art by those skilled in the art without departing from the scope of the subject art. For example, a different number of a given module or unit may be used, or one or more different types of a given module or unit may be used, or a given module or unit may be added, or a given module or unit may be omitted.

[0163] Some versions of the embodiments described herein may be implemented using a processor that is part of a computer system and capable of communicating with several peripheral devices via a bus subsystem. Versions of the embodiments described herein implemented using a computer system may be implemented using a general-purpose computer programmed to perform the methods described herein. Alternatively, versions of the embodiments described herein implemented using a computer system may be implemented using a purpose-specific computer built with hardware configured to perform the methods described herein. Versions of the embodiments described herein may also be implemented using a combination of at least one general-purpose computer and at least one purpose-specific computer.

[0164] In versions implemented using computer systems, each processor may include the computer system's central processing unit (CPU), microprocessor, application-specific integrated circuit (ASIC), other types of hardware components, and combinations thereof. A computer system may include multiple types of processors. Peripherals of a computer system may include, for example, a storage subsystem including memory devices and file storage subsystems, user interface input devices, user interface output devices, and a network interface subsystem. Input and output devices may enable 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 another computer system. User interface input devices may include keyboards; pointing devices such as mice, trackballs, touchpads, or graphics tablets; scanners; touchscreens integrated into displays; voice input devices such as speech recognition systems and microphones; and other types of input devices. In general, the use of the term “input device” is intended to encompass all possible types of devices and methods for inputting information into a computer system.

[0165] In versions implemented using computer systems, user interface output devices may include non-visual displays such as display subsystems, printers, facsimile machines, or audio output devices. Display subsystems may include flat-panel devices such as cathode ray tubes (CRTs), liquid crystal displays (LCDs), projection devices, or any other mechanism for generating visible images. Display subsystems may also provide non-visual displays such as audio output devices. Generally, the use of the term “output device” is intended to include all possible types of devices and methods for outputting information from a computer system to a user or another machine or computer system.

[0166] In versions implemented using a computer system, the storage subsystem can store programming and data structures that provide some or all of the functionality of the modules and methods described herein. These software modules can generally be executed by the computer system's processor alone or in combination with other processors. The memory used by the storage subsystem may include multiple memories, including main random access memory (RAM) for storing instructions and data during program execution, and read-only memory (ROM) for storing fixed instructions. The file storage subsystem can provide persistent storage for program and data files and may include a hard disk drive, a floppy disk drive with associated removable media, a CD-ROM drive, an optical drive, or a removable media cartridge. Modules implementing the functionality of a particular embodiment may be stored by the file storage subsystem within the storage subsystem or in another machine accessible to the processor.

[0167] In versions implemented using computer systems, the computer system itself may be of various types, including personal computers, portable computers, workstations, computer terminals, network computers, televisions, mainframes, server farms, widely-distributed sets of loosely networked computers, or any other data processing systems or user devices. Due to the constantly changing nature of computers and networks, the examples of computer systems described herein are intended only to be concrete examples illustrating the disclosed technology. Many other configurations of computer systems are possible, having more or fewer components than those described herein.

[0168] As a product rather than a method, a non-temporary computer-readable medium (CRM) can be loaded with program instructions executable by a processor. When executed, these program instructions perform one or more of the computer implementation methods described above. Alternatively, the program instructions, loaded onto a non-temporary CRM and combined with appropriate hardware, can become one or more components of a computer implementation system that performs the disclosed methods.

[0169] Underlined and / or italicized headings and subheadings are used for convenience only and are not intended to limit the scope of the subject art, nor are they to be referenced in connection with the interpretation of the description of the subject art. All structural and functional equivalents of elements of the various embodiments described throughout this disclosure, whether known to those skilled in the art or to become known later, are expressly incorporated herein by reference and are intended to be included by the subject art. Furthermore, nothing disclosed herein is intended to be made available to the public, whether such disclosure is expressly stated in the above description or not.

[0170] It should be recognized that all combinations of the aforementioned concepts and any additional concepts discussed in further detail below (insofar as such concepts do not contradict each other) are intended to be part of the subject matter of the inventions disclosed herein. In particular, all combinations of the claimed subject matter listed at the end of this disclosure are intended to be part of the subject matter of the inventions disclosed herein.

[0171] [Implementation Method] (1) A microfluidic device, (a) The first plate and (b) The second plate, (c) A microfluidic path defined between the first plate and the second plate, wherein the microfluidic path includes at least one chamber, (d) An elastic layer disposed between the first plate and the second plate, wherein the elastic layer is (i) A first membrane extending across at least one chamber, wherein the first membrane is configured to deflect to drive a fluid through the at least one chamber, (ii) A second membrane having at least one opening, wherein the at least one opening is aligned with the at least one chamber, An elastic layer including, Microfluidic devices, including those mentioned above. (2) The microfluidic apparatus according to Embodiment 1, wherein the second membrane does not extend across the at least one chamber. (3) The microfluidic apparatus according to any one of embodiments 1 to 2, wherein the at least one chamber extends along a central longitudinal axis, and the at least one opening has a center point defined with respect to the central longitudinal axis. (4) The microfluidic apparatus according to any one of embodiments 1 to 3, wherein the first membrane is configured to fluidly isolate a first region of the at least one chamber from a second region of the at least one chamber. (5) The microfluidic apparatus according to Embodiment 4, wherein the at least one opening is located directly above the first region and directly below the second region.

[0172] (6) A microfluidic apparatus according to any one of embodiments 1 to 5, wherein the first membrane and the second membrane are stacked on top of each other. (7) The microfluidic apparatus according to any one of embodiments 1 to 6, wherein one of the first membrane or the second membrane is bonded to the first plate, and the other of the first membrane or the second membrane is bonded to the second plate. (8) A microfluidic apparatus according to any one of embodiments 1 to 7, wherein at least one of the first or second membrane comprises polydimethylsilicone (PDMS). (9) A microfluidic apparatus according to any one of embodiments 1 to 8, wherein at least one of the first or second membrane has a thickness of about 100 μm. (10) A microfluidic device according to any one of embodiments 1 to 9, wherein the elastic layer has an overall thickness of about 200 μm.

[0173] (11) The microfluidic apparatus according to any one of embodiments 1 to 10, wherein the at least one chamber and the at least one opening have the same profile as each other. (12) The microfluidic apparatus according to Embodiment 11, wherein the profile is at least one of circular, oval, or at least partially rectangular. (13) The microfluidic device according to any one of embodiments 1 to 12, wherein the at least one chamber includes a valve chamber. (14) The microfluidic apparatus according to any one of embodiments 1 to 13, wherein the at least one chamber comprises a plurality of chambers, and the at least one opening comprises a plurality of openings, and each of the plurality of openings is aligned with a corresponding chamber of the plurality of chambers. (15) The microfluidic apparatus according to Embodiment 14, wherein the plurality of openings have different profiles from each other.

[0174] (16) A microfluidic device, (a) Upper plate and (b) Lower plate and (c) A microfluidic path defined between the upper plate and the lower plate, wherein the microfluidic path includes a chamber, (d) An elastic layer disposed between the upper plate and the lower plate, wherein the elastic layer is (i) A first membrane that bisects the chamber so as to define an upper chamber region and a lower chamber region, wherein the first membrane is configured to deflect in order to drive a fluid through the chamber, (ii) A second membrane having an opening, the opening being located directly below the upper chamber region and directly above the lower chamber region, An elastic layer including, Microfluidic devices, including those mentioned above. (17) The microfluidic apparatus according to Embodiment 16, wherein the first membrane and the second membrane are stacked on top of each other. (18) A microfluidic device according to any one of embodiments 16 to 17, wherein the first membrane is located below the second membrane. (19) A microfluidic apparatus according to any one of embodiments 16 to 18, wherein the chamber and the opening have the same profile as each other. (20) A microfluidic device, (a) The first plate and (b) The second plate, (c) A microfluidic path defined between the first plate and the second plate, wherein the microfluidic path includes a chamber, (d) An elastic layer disposed between the first plate and the second plate, wherein the elastic layer is (i) A first membrane configured to fluidly isolate a first region of the chamber from a second region of the chamber, wherein the first membrane is configured to deflect to drive fluid through the chamber, (ii) A second membrane having an opening, the opening being aligned with the chamber, An elastic layer including, Microfluidic devices, including those mentioned above.

[0175] (21) A microfluidic device, (a) The first plate and (b) The second plate, (c) An elastic layer disposed between the first plate and the second plate, (d) A microfluidic path defined between the first plate and the second plate, the microfluidic path including a chamber, the chamber extending along a central longitudinal axis, and at least a portion of the chamber having a flat bell-shaped cross-sectional profile, Microfluidic devices, including those mentioned above. (22) The microfluidic apparatus according to Embodiment 21, wherein the elastic layer extends across the chamber to define a first region and a second region of the chamber, and at least one of the first region or the second region has the flat bell-shaped cross-sectional profile. (23) The microfluidic apparatus according to embodiment 22, wherein each of the first region and the second region has the flat bell-shaped cross-sectional profile. (24) The microfluidic apparatus according to any one of embodiments 21 to 23, wherein the chamber is substantially symmetrical with respect to the central longitudinal axis. (25) The microfluidic apparatus according to any one of embodiments 21 to 24, wherein the chamber includes a radially inner surface, a radially outer surface, and a radially intermediate surface that collectively define the flat bell-shaped cross-sectional profile.

[0176] (26) The microfluidic device according to embodiment 25, wherein the radially inner surface has a flat cross-sectional profile. (27) The microfluidic apparatus according to any one of embodiments 25 to 26, wherein the radially outer surface and the radially intermediate surface each have a curved cross-sectional profile. (28) The microfluidic device according to embodiment 27, wherein the radially outer surface has a convex cross-sectional profile. (29) The microfluidic apparatus according to any one of embodiments 27 to 28, wherein the radial intermediate surface has a concave cross-sectional profile. (30) The microfluidic apparatus according to any one of embodiments 27 to 29, wherein the radially outer surface and the radially intermediate surface collectively define at least one of an S-shaped cross-sectional profile or an inverted S-shaped cross-sectional profile.

[0177] (31) The microfluidic apparatus according to any one of embodiments 25 to 30, wherein the radially inner surface is generally circular. (32) The microfluidic apparatus according to embodiment 31, wherein the radially outer surface and the radially intermediate surface are each generally annular. (33) A microfluidic apparatus according to any one of embodiments 25 to 32, wherein a portion of the elastic layer is configured to deflect to drive a fluid through the chamber, and the portion of the elastic layer is configured to conform to the radially inner surface, the radially outer surface, and the radially intermediate surface, respectively, when deflected. (34) The elastic layer is (i) A first membrane extending across the chamber, the first membrane being configured to deflect to drive the fluid through the chamber, and the first membrane being configured to conform to the radially inner surface, the radially outer surface, and the radially intermediate surface when deflected, (ii) A second membrane having an opening, the opening being aligned with the chamber, A microfluidic apparatus according to embodiment 33, including the above. (35) The microfluidic device according to any one of embodiments 21 to 34, wherein the chamber includes a valve chamber.

[0178] (36) A microfluidic device, (a) The first plate and (b) The second plate, (c) An elastic layer disposed between the first plate and the second plate, (d) A microfluidic path defined between the first plate and the second plate, the microfluidic path including a chamber, the chamber extending along a central longitudinal axis, (i) A radially inner surface having a flat cross-sectional profile, (ii) A radial outer surface having a convex cross-sectional profile, (iii) A radial intermediate surface having a concave cross-sectional profile, Microfluidic pathways, Microfluidic devices, including those mentioned above. (37) The microfluidic apparatus according to embodiment 36, wherein the radially outer surface and the radially intermediate surface collectively define at least one of an S-shaped cross-sectional profile or an inverted S-shaped cross-sectional profile. (38) The microfluidic device according to any one of embodiments 36 to 37, wherein the radially inner surface is generally circular. (39) The microfluidic apparatus according to embodiment 38, wherein the radially outer surface and the radially intermediate surface are each generally annular. (40) A microfluidic device, (a) The first plate and (b) The second plate, (c) An elastic layer disposed between the first plate and the second plate, (d) A microfluidic path defined between the first plate and the second plate, the microfluidic path including a chamber, the chamber extending along a central longitudinal axis, (i) radially inner surface and (ii) Radial outer surface and (iii) Radial intermediate surface and Includes, (A) The radially outer surface extends convexly from the radially outer circumference of the chamber to the radially inward on the first side of the longitudinal axis, (B) The radial intermediate surface extends concavely inward from the radially inner end of the radially outer surface on the first side of the longitudinal axis, (C) The radially inner surface extends from the radially inner end of the radially intermediate surface across the longitudinal axis to the second side of the longitudinal axis, substantially parallel to the first plate and the second plate. (D) The radial intermediate surface extends concavely outward from the end of the radial inner surface on the second side of the longitudinal axis, (E) The radially outer surface extends convexly outward from the radially outer end of the radially intermediate surface on the second side of the longitudinal axis, Microfluidic pathways and Microfluidic devices, including those mentioned above.

Claims

1. A microfluidic device, (a) The first plate and (b) The second plate and (c) A microfluidic path defined between the first plate and the second plate, wherein the microfluidic path includes at least one chamber, (d) An elastic layer disposed between the first plate and the second plate, wherein the elastic layer is (i) A first membrane extending across the at least one chamber, the first membrane being configured to deflect to drive a fluid through the at least one chamber, (ii) A second membrane having at least one opening, wherein the at least one opening is aligned with the at least one chamber, An elastic layer including, Microfluidic devices, including those mentioned above.

2. The microfluidic apparatus according to claim 1, wherein the second membrane does not extend across the at least one chamber.

3. The microfluidic apparatus according to claim 1, wherein the at least one chamber extends along a central longitudinal axis, and the at least one opening has a central point defined with respect to the central longitudinal axis.

4. The microfluidic apparatus according to claim 1, wherein the first membrane is configured to fluidly isolate a first region of the at least one chamber from a second region of the at least one chamber.

5. The microfluidic apparatus according to claim 4, wherein the at least one opening is located directly above the first region and directly below the second region.

6. The microfluidic apparatus according to claim 1, wherein the first membrane and the second membrane are stacked on top of each other.

7. The microfluidic apparatus according to claim 1, wherein one of the first or second membrane is bonded to the first plate, and the other of the first or second membrane is bonded to the second plate.

8. The microfluidic apparatus according to claim 1, wherein at least one of the first or second membrane comprises polydimethylsilicone (PDMS).

9. The microfluidic apparatus according to claim 1, wherein at least one of the first or second membrane has a thickness of about 100 μm.

10. The microfluidic apparatus according to claim 1, wherein the elastic layer has an overall thickness of about 200 μm.

11. The microfluidic apparatus according to claim 1, wherein the at least one chamber and the at least one opening have the same profile as each other.

12. The microfluidic apparatus according to claim 11, wherein the profile is at least one of circular, oval, or at least partially rectangular.

13. The microfluidic apparatus according to claim 1, wherein the at least one chamber includes a valve chamber.

14. The microfluidic apparatus according to claim 1, wherein the at least one chamber comprises a plurality of chambers, and the at least one opening comprises a plurality of openings, and each of the plurality of openings is aligned with a corresponding chamber among the plurality of chambers.

15. The microfluidic apparatus according to claim 14, wherein the plurality of openings have different profiles from each other.

16. A microfluidic device, (a) Upper plate and (b) Lower plate and (c) A microfluidic path defined between the upper plate and the lower plate, wherein the microfluidic path includes a chamber, (d) An elastic layer disposed between the upper plate and the lower plate, wherein the elastic layer is (i) A first membrane that bisects the chamber so as to define an upper chamber region and a lower chamber region, wherein the first membrane is configured to deflect in order to drive a fluid through the chamber, (ii) A second membrane having an opening, wherein the opening is located directly below the upper chamber region and directly above the lower chamber region, An elastic layer including, Microfluidic devices, including those mentioned above.

17. The microfluidic apparatus according to claim 16, wherein the first film and the second film are laminated together.

18. The microfluidic apparatus according to claim 16, wherein the first membrane is located below the second membrane.

19. The microfluidic apparatus according to claim 16, wherein the chamber and the opening have the same profile as each other.

20. A microfluidic device, (a) The first plate and (b) The second plate and (c) A microfluidic path defined between the first plate and the second plate, wherein the microfluidic path includes a chamber, (d) An elastic layer disposed between the first plate and the second plate, wherein the elastic layer is (i) A first membrane configured to fluidly isolate a first region of the chamber from a second region of the chamber, wherein the first membrane is configured to deflect to drive a fluid through the chamber, (ii) A second membrane having an opening, the opening being aligned with the chamber, An elastic layer including, Microfluidic devices, including those mentioned above.

21. A microfluidic device, (a) The first plate and (b) The second plate and (c) An elastic layer disposed between the first plate and the second plate, (d) A microfluidic path defined between the first plate and the second plate, the microfluidic path including a chamber, the chamber extending along a central longitudinal axis, (i) A radially inner surface having a flat cross-sectional profile, (ii) A radially outer surface having a convex cross-sectional profile, (iii) A radial intermediate surface having a concave cross-sectional profile, Microfluidic pathways, Microfluidic devices, including those mentioned above.

22. The microfluidic apparatus according to claim 21, wherein the radially outer surface and the radially intermediate surface collectively define at least one of an S-shaped cross-sectional profile or an inverted S-shaped cross-sectional profile.

23. The microfluidic apparatus according to claim 21, wherein the radially inner surface is generally circular.

24. The microfluidic apparatus according to claim 23, wherein the radially outer surface and the radially intermediate surface are each generally annular.

25. A microfluidic device, (a) The first plate and (b) The second plate and (c) An elastic layer disposed between the first plate and the second plate, (d) A microfluidic path defined between the first plate and the second plate, the microfluidic path including a chamber, the chamber extending along a central longitudinal axis, (i) radially inner surface and (ii) Radially outer surface and (iii) Radial intermediate surface and Includes, (A) The radially outer surface extends convexly from the radially outer circumference of the chamber to the radially inward on the first side of the longitudinal axis, (B) The radial intermediate surface extends in a concave shape radially inward from the radially inner end of the radially outer surface on the first side of the longitudinal axis, (C) The radially inner surface extends from the radially inner end of the radially intermediate surface across the longitudinal axis to the second side of the longitudinal axis, generally parallel to the first plate and the second plate. (D) The radial intermediate surface extends concavely outward from the end of the radial inner surface on the second side of the longitudinal axis, (E) The radially outer surface extends convexly outward from the radially outer end of the radially intermediate surface on the second side of the longitudinal axis, Microfluidic pathways and Microfluidic devices, including those mentioned above.

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