System and method for multi-angle detection of dynamic light scattering

Abstract

This specification describes a system and method for acquiring dynamic light scattering data from a plurality of particle samples and, in some cases, also measuring the UV / Vis absorption spectrum of the particle samples. The system and method can characterize particles within a sample by acquiring light scattering data from multiple angles using a single rotating photodetector. Data acquired in this manner can allow for a larger dynamic range of particle sizing than when using a single angle.

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 618,653 filed on January 8, 2024, the entirety of which is incorporated by reference.

[0003] field

[0004] This application relates to a system and method for characterizing particles, e.g., nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. Characterization may include acquiring light scattering and / or UV / Vis data associated with the particles. When light scattering data is acquired, it may be detected using a rotation detector capable of measuring light scattered from multiple angles. Background Technology

[0005] Dynamic light scattering is a commonly known method for analyzing particles in which measurements of scattered light over time are used to determine the particle size or size distribution. Particle characteristics can be inferred from the temporal variation of the scattered light. For example, autocorrelation can be performed on the time series of scattered light intensity, followed by a fit to the autocorrelation function (e.g., Cumulants, CONTIN, NNLS / non-negative least squares) to determine particle characteristics. In other cases, the Fourier transform can be used to determine the power spectrum of the scattered light, and a similar fit to the power spectrum can be used to determine particle characteristics.

[0006] Generally, when performing dynamic light scattering measurements, light scattered at a single, well-defined angle is used. However, the direction and number of scattered photons depend on the particle size. For example, if the particles are small, light can be scattered fairly uniformly in all directions. If the particles are large, more light may be scattered forward than backward. Therefore, when analyzing a mixture of particle sizes (e.g., some small and some large), some particles may not be visible at all, or light scattering results may be inaccurate when measured at a single angle, because each particle size generates a unique scattering pattern. In other words, particle-related results can vary depending on the angle used to detect the scattered light.

[0007] Accordingly, to acquire dynamic light scattering measurements for a sample, a method for detecting scattered light from more than one angle has been developed. Some conventional systems acquire multi-angle measurements by positioning multiple photodetectors or multiple photoemitters around a single sample of interest. However, these systems have several drawbacks, including, for example, that processing the results is time-consuming due to a limited range of detection angles for scattered light and / or the need to match information between multiple photodetectors and multiple photoemitters.

[0008] Therefore, there is a need for a new and useful system for detecting light scattered from multiple angles. A method for determining particle size and / or particle size distribution within a sample may also be useful.

[0009] This specification describes a system and method for acquiring dynamic light scattering data from a particle sample and, in some cases, also measuring the UV / Vis absorption spectrum of the particle sample, e.g., a microliter-sized sample. The system and method can characterize particles within a sample by acquiring light scattering data from multiple angles. Data acquired in this manner may allow for a larger dynamic range of particle sizing than when using a single angle. Additionally, quantitative data on how many of each particle size are present within the sample can be acquired. The system and method can combine absolute correlation functions at each angle to provide a single result independent of the angle, which can help to measure previously hidden particle size populations (e.g., due to weak scattering) and / or generate a more complete size distribution. In some variations, dynamic light scattering measurements may be acquired using a system comprising a single rotating light detector, e.g., a single rotating light fiber. A single photodetector can detect light scattered at multiple angles by rotating on a sample plate that holds multiple samples for particle characterization.

[0010] A system for characterizing particles may include one or more processors, a plurality of samples, and a first module configured to measure light scattered by each of the plurality of samples. Each of the plurality of samples may include one or more types of particles. A sample may include any type of particle that scatters light when illuminated. Exemplary types of particles may include, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof.

[0011] In one variation, the first module may include one or more light emitters, an actuator having a rotation axis, and a single photodetector configured to rotate about the rotation axis and receive light scattered by each sample at a plurality of scattering angles. One or more light emitters may include a laser light source. In some variations, one or more light emitters of the first module of the system may include two light emitters, namely a first light emitter and a second light emitter. The first light emitter may be placed above the sample plate, and the second light emitter may be placed below the sample plate. It is understood that the light emitters may be configured in various other ways.

[0012] Multiple samples may be held on a sample plate of the system. More specifically, multiple samples may be held in multiple cuvettes on the sample plate. In some variations, each of the multiple samples on the sample plate may contain the same type of particle. In other variations, each of the multiple samples on the sample plate may contain a different type of particle. In further variations, each of the multiple samples may contain a mixture of particles. In further variations, some samples on the sample plate may contain the same particle or a mixture of the same particle, and other samples on the plate may contain different types of particles or a mixture of different particles.

[0013] The first module may also be configured to adjust the position of the emission beam axis so that the intersection [beam overlap] between the emission beam axis and the detector beam axis is restored during each rotation of the photodetector and / or when the illumination direction changes (e.g., when illumination from the top of the sample plate changes to illumination from the bottom of the sample plate and vice versa). The first module may be configured so that the position of the emission beam axis can be adjusted automatically or manually. Additionally, the first module may be configured to include a mechanism for maintaining the beam overlap at the same height across multiple cuvettes of the sample plate. Some variations of the system may include a second module configured to automatically adjust the position of the sample plate according to instructions from one or more processors.

[0014] Instead of multiple photodetectors, the system described herein generally comprises a single photodetector. The single photodetector may be configured to rotate about 1.0 degree to about 280 degrees around the axis of rotation of a rotating actuator, including all values ​​and sub-ranges therein. The single photodetector may include an optical fiber.

[0015] A single photodetector may be configured to receive forward light scattering from multiple scattering angles in the range of about 25 degrees to about 45 degrees, including all values ​​and sub-ranges therein. With respect to backward light scattering, a single photodetector may be configured to receive backward light scattering from multiple scattering angles in the range of about 105 degrees to about 170 degrees, including all values ​​and sub-ranges therein.

[0016] Some variations of the system described herein may also be configured to acquire UV / Vis data from a plurality of samples. In such variations, the system may include a third module configured to illuminate a plurality of samples with light having a wavelength in the range of about 190 nm to about 900 nm and to measure the absorbance of the plurality of samples. The particles in these samples may be those mentioned above, e.g., nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules.

[0017] The volume of the sample analyzed by the system, including all values ​​and sub-ranges therein, can be significantly small, ranging from about 0.5 μl to about 2.5 μl. In one variation, the volume of multiple samples is about 2 μl. In another variation, the volume of multiple samples is about 2 μl or less.

[0018] The sample plate may have various sizes. For example, the sample plate may have a length ranging from about 11 cm to about 13 cm (including all values ​​and sub-ranges within it) and a width ranging from about 7.0 cm to about 9.0 cm (including all values ​​and sub-ranges within it). In one instance, the sample plate may have a length of about 12.8 cm and a width of about 8.5 cm.

[0019] The sample plate may also be configured in various ways. In some variations, the sample plate may comprise a plurality of members, wherein each member of the plurality of members may be configured as a strip having a length and a width (referred herein to as a “chip” or “microfluidic chip”). The length of the strip may be in the range of about 7.0 cm to about 9.0 cm (including all values ​​and sub-ranges therein), and the width of the strip may be in the range of about 1.0 cm to about 3.0 cm (including all values ​​and sub-ranges therein). For example, it may be useful for the strip to have a length of about 8.1 cm and a width of about 1.8 cm. Any suitable number of strips may be included on the sample plate, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In one variation, the sample plate may include 6 strips.

[0020] Each strip may include a plurality of wells configured to receive a sample. Each well of the plurality of wells may be coupled to a corresponding cuvette (of the plurality of cuvettes) by a channel, e.g., a microchannel. The cuvette may have a path length ranging from about 0.1 mm to about 0.7 mm, including all values ​​and sub-ranges within it. In some variations, the path length of the cuvette is about 0.1 mm. In other variations, the path length of the cuvette is about 0.7 mm.

[0021] One or more processors of the system may be configured to determine the size of one or more types of particles in a sample based on data from scattered light. The size of one or more types of particles in a sample may be in the range of about 0.3 nm to about 3,000 nm, including all values ​​and sub-ranges therein. For example, the size of one or more particles is approximately 0.3 nm, approximately 0.4 nm, approximately 0.5 nm, approximately 0.6 nm, approximately 0.7 nm, approximately 0.8 nm, approximately 0.9 nm, approximately 1.0 nm, approximately 1.5 nm, approximately 2.0 nm, approximately 2.5 nm, approximately 3.0 nm, approximately 3.5 nm, approximately 4.0 nm, approximately 4.5 nm, approximately 5.0 nm, approximately 5.5 nm, approximately 6.0 nm, approximately 6.5 nm, approximately 7.0 nm, approximately 7.5 nm, approximately 8.0 nm, approximately 8.5 nm, approximately 9.0 nm, approximately 9.5 nm, approximately 10 nm, approximately 15 nm, approximately 20 nm, approximately 25 nm, approximately 30 nm, approximately 35 nm, approximately 40 nm, approximately 45 nm, approximately 50 nm, approximately 55 nm, approximately It may be 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 1,000 nm, about 1,500 nm, about 2,000 nm, about 2,500 nm, or about 3,000 nm. One or more processors may be configured to receive multiple signals from a single photodetector, wherein each of the multiple signals may represent scattered light at a specific angle.Next, data from scattered light at multiple (e.g., different) angles can be combined and analyzed by one or more processors to determine a single result that may be independent of the angle. In some variations, one or more processors may utilize an algorithm configured to determine particle size using data and / or signals related to one or more of sample temperature, sample viscosity, hydrodynamic diameter of the particle, intensity of scattered light, ratio of scattered light intensity to incident light intensity at a specific angle, angle of scattered light, and rate of change of scattered light intensity (e.g., variation of scattered light intensity over time). One or more processors may include a correlator configured to analyze particle size based on the data and / or intensity of the various signals described above.

[0022] The present specification also describes a method for characterizing particles. Exemplary particles include, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. Generally, the method may include the steps of illuminating a plurality of samples with light from one or more light emitters—wherein each of the plurality of samples comprises one or more types of particles—; rotating a photodetector over a plurality of scattering angles around a rotation axis; detecting light scattered by particles over a plurality of scattering angles; and obtaining light scattering measurements for each of the plurality of scattering angles. As mentioned above, data from the scattered light at the plurality of scattering angles may then be combined and analyzed by one or more processors to determine a single result that may be independent of the angle.

[0023] One or more light emitters may include a laser light source. It is understood that suitable alternative light sources may also be used. Some variations of the method may utilize a first light emitter and a second light emitter. In such variations, the first light emitter may be placed above a plurality of samples, and the second light emitter may be placed below a plurality of samples.

[0024] In some variations, the method may further include a step of adjusting the position of the emission beam axis so that the intersection point between the emission beam axis and the detector axis is maintained at the same height in a plurality of cuvettes contained in the sample plate. The position of the emission beam axis may be adjusted manually or automatically upon instruction from one or more processors. Similarly, the position of the sample plate may be adjusted manually or automatically.

[0025] Automatic adjustment of the emission beam axis may be based on instructions from one or more processors, including one or more preliminary positioning steps. For example, one or more preliminary positioning steps may include a step of moving one or more light emitters to the smallest circle on a hyperboloid. In some variations, the method may further include one or more fine positioning steps, such as determining the maximum intensity of light scattered by each sample at a plurality of scattering angles.

[0026] When the photodetector is rotated to acquire scattered light at multiple angles, the rotation angle may be in the range of about 1 degree to about 280 degrees, including all values ​​and sub-ranges within it, around the rotation axis of the actuator. Forward light scattering may be detected at multiple scattering angles in the range of about 25 degrees to about 45 degrees, including all values ​​and sub-ranges within it, and backward light scattering may be detected at multiple scattering angles in the range of about 105 degrees to about 170 degrees, including all values ​​and sub-ranges within it.

[0027] Additionally, the method may include the step of acquiring UV / Vis data from a plurality of samples. For example, the plurality of samples may be illuminated with light having a wavelength in the range of about 190 nm to about 900 nm, and the absorbance may be measured.

[0028] Characterization of particles can be achieved for small sample volumes ranging from about 0.5 μl to about 2.5 μl, including all values ​​and sub-ranges within them. In one variation, the volume of multiple samples is about 2 μl. In another variation, the volume of multiple samples is about 2 μl or less.

[0029] Determining the size of one or more types of particles within a sample can be achieved by one or more processors of a system configured to analyze data from scattered light. The size of one or more types of particles within a sample, as described above, may be in the range of about 0.3 nm to about 3,000 nm, including all values ​​and sub-ranges therein. Once a signal is acquired from scattered light detected by a single photodetector at multiple scattering angles, the data from scattered light at multiple (e.g., different) angles may then be combined and analyzed by one or more processors to determine a single result (e.g., a single solution). The single result may be independent of angles. In some variations, one or more processors may determine the size of one or more types of particles within each sample based on the shape of the absolute correlation function of multiple scattering angles. Brief explanation of the drawing

[0030] The patent or application file includes at least one drawing in color. A copy of such patent or patent application publication using the color drawing(s) will be provided by the Patent Office upon request and payment of the necessary fees. FIG. 1a depicts a front view of the housing of an exemplary system for measuring light scattering at multiple angles. FIG. 1b depicts a module inside the housing shown in FIG. 1a. FIG. 2a depicts a perspective view of an exemplary first module (dynamic light scattering module) of a system for measuring light scattering at multiple angles. FIGS. 2B and 2C depict an exemplary actuator configured to rotate a photodetector so that scattered light can be detected at multiple scattering angles. Figures 3a and 3b depict an example of beam overlap between the emitted beam and the receiving beam when the photodetector rotates. In Figure 3a, the beam overlap is restored. In Figure 3b, the beam overlap is not restored. FIG. 3c illustrates an exemplary first module comprising a stage configured to move one or more light emitters in a horizontal plane in the X' and / or Y' directions so that beam overlap can be restored. FIG. 4a depicts an exemplary first module comprising an actuator configured to translate the entire first module up and down so that beam overlap can occur at the same height in the cuvette(s). FIGS. 4b and FIGS. 4c illustrate beam overlap at the same height in the cuvette when the cuvette is illuminated from the top of the sample plate (Fig. 4b) and when it is illuminated from the bottom of the sample plate (Fig. 4c). Figure 5a depicts an exemplary sample plate. FIG. 5b shows one of the multiple strips provided on the sample plate of FIG. 5a. Fig. 5c is an enlarged view of the structure of the strip shown in Fig. 5b. Figures 6a through 6d depict an exemplary sample characterization cycle of the system. FIGS. 7a through 7d illustrate an exemplary algorithm for restoring beam overlap. FIGS. 8a through 8c depict exemplary 2D representations of an absolute correlation function whose shape can help determine the size of one or more types of particles within a sample. Specific details for implementing the invention

[0031] This specification describes a system and method for acquiring dynamic light scattering data from a particle sample and, in some cases, also measuring the UV / Vis absorption spectrum of the particle sample. The system and method can characterize particles within a sample by acquiring light scattering data from multiple angles using a single photodetector that rotates due to the rotation of a coupled actuator. Data acquired in this manner may allow for a larger dynamic range of particle size determination than when using a single angle. The system and method can provide a single result independent of the angle by combining absolute correlation functions at each angle. Additionally, the system and method may be designed for manual workflows, automated workflows, or workflows of both types (e.g., switching between manual and automated workflows and vice versa).

[0032] System

[0033] The system described herein may include various modules capable of assisting in characterizing particles within one or more samples, either individually or collectively. Generally, the system may be sized and / or shaped to be portable and / or placed on a workbench, table, shelf, etc. In some variations, the system may include at least a first module, a second module, and a third module contained within a housing, and these modules may be configured to characterize the particle sample and / or position the sample for characterization.

[0034] For example, as illustrated in FIGS. 1a and 1b, the system (100) may include a housing (102), and within the housing (102), a first module (104), a second module (106), and a third module (108) may exist on a base plate (116). A plate drawer of the second module (106), such as a microplate drawer (110), may be configured to open and extend through the front wall (112) of the housing (102) to an open position so that a sample plate (114) can be inserted into the drawer (110), and may be configured to retract back to a closed position within the housing (102) to place the sample plate (114) within the housing (102). Once placed within the housing (102), a plurality of samples within the sample plate (114) may be analyzed by one or more of the modules of the system (100). A multi-indicator light (111) may also be provided on the front wall (112) of the housing (102) to indicate the status of the system (e.g., various statuses). For example, a flashing white light may indicate that the system is booting; a lit white light may indicate that the system has booted normally and is ready for use; a flashing blue light may indicate that the microplate drawer is moving in / out or is open, that the plate is still inside the system, and / or that the system is still measuring; a lit red light may indicate that the system is in an error state; a flashing red light may indicate that the microplate drawer is obstructed; and a flashing yellow light may indicate a self-test mode after startup.

[0035] One module of the system may be combined with one or more different modules of the system via mechanical connection (e.g., by cable) or wireless connection. Components of the system, such as housings, modules, and sample plates, may be made of any suitable material, such as any suitable polymer material, metal material, or glass material, or any combination of suitable materials.

[0036] Module 1 (Dynamic Light Scattering Module)

[0037] As previously described, a system for characterizing particles may include one or more processors, a plurality of samples, and a first module configured to measure light scattered by each of the plurality of samples. One or more processors may be coupled to the first module (and / or other modules, e.g., a second module or a third module) by a wired or wireless connection. Each of the plurality of samples may contain one or more types of particles. The samples may contain any type of particle that scatters light when illuminated. Exemplary types of particles may include, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof.

[0038] The first module may include one or more light emitters, a single photodetector, and an actuator having a rotation axis. One or more light emitters may include a source of laser light. The laser light source may be any laser source or configuration capable of providing monochromatic light and / or polarization. In some variations, the laser source may have an output of about 40 mW and may produce a wavelength of about 660 nm. When multiple light emitters are used, the multiple light emitters may include the same or different laser sources. The laser source(s) may be contained within the housing of the system.

[0039] A single photodetector may include an optical fiber and may be coupled to an actuator. Coupling between the photodetector and the actuator may be achieved using a clamp, screw or key connection, or other types of connection. In some variations, the single photodetector may be coupled to the actuator by a clamp such that rotation of the actuator, in turn, rotates the photodetector around the actuator's axis of rotation. Rotation of the single photodetector around the axis of rotation may allow the photodetector to receive light scattered by each sample at multiple scattering angles. The single photodetector may rotate over a sample plate holding multiple samples to detect light scattered at multiple scattering angles.

[0040] In some variations, a single photodetector can rotate about 1.0 degree to about 280 degrees around a rotation axis, including all values ​​and sub-ranges within it. In other words, the angle sweep of a single photodetector can be in the range of about 1.0 degree to about 280 degrees, including all values ​​and sub-ranges within it. For example, a single photodetector rotates about 1.0°, 5.0°, 10°, 15°, 20°, 25°, 30°, 35°, 40°, 45°, 50°, 55°, 60°, 65°, 70°, 75°, 80°, 85°, 90°, 95°, 100°, 105°, 110°, 115°, 120°, 125°, 130°, 135°, 140°, 145°, 150°, 155°, 160°, 165°, 170°, 175°, and approximately around the axis of rotation. It can rotate 180 degrees, approximately 185 degrees, approximately 190 degrees, approximately 195 degrees, approximately 200 degrees, approximately 205 degrees, approximately 210 degrees, approximately 215 degrees, approximately 220 degrees, approximately 225 degrees, approximately 250 degrees, approximately 255 degrees, approximately 260 degrees, approximately 265 degrees, approximately 270 degrees, approximately 275 degrees, or approximately 280 degrees. In some variations, a single photodetector can rotate 280 degrees around the axis of rotation or more than 280 degrees around the axis of rotation. In other variations, rotation of the actuator between approximately 30 degrees and approximately 180 degrees also rotates the single photodetector by a corresponding angle.

[0041] The scattered light received by a single light detector may include forward scattered light and / or backscattered light from a plurality of scattering angles. When forward light scattering is received, the plurality of scattering angles may be in the range of about 25 degrees to about 45 degrees, including all values ​​and sub-ranges therein. For example, the forward light scattering angles of the plurality of scattering angles may be about 25 degrees, about 26 degrees, about 27 degrees, about 28 degrees, about 29 degrees, about 30 degrees, about 31 degrees, about 32 degrees, about 33 degrees, about 34 degrees, about 35 degrees, about 36 degrees, about 37 degrees, about 38 degrees, about 39 degrees, about 40 degrees, about 41 degrees, about 42 degrees, about 43 degrees, about 44 degrees, or about 45 degrees.

[0042] When backlight scattering is received, multiple scattering angles may be in the range of about 105 degrees to about 170 degrees, including all values ​​and sub-ranges within them. For example, the backlight scattering angles of multiple scattering angles are approximately 105°, approximately 106°, approximately 107°, approximately 108°, approximately 109°, approximately 110°, approximately 111°, approximately 112°, approximately 113°, approximately 114°, approximately 115°, approximately 116°, approximately 117°, approximately 118°, approximately 119°, approximately 120°, approximately 121°, approximately 122°, approximately 123°, approximately 124°, approximately 125°, approximately 126°, approximately 127°, approximately 128°, approximately 129°, approximately 130°, approximately 131°, approximately 132°, approximately 133°, approximately 134°, approximately 135°, approximately 136°, approximately 137°, approximately It may be 138 degrees, approximately 139 degrees, approximately 140 degrees, approximately 141 degrees, approximately 142 degrees, approximately 143 degrees, approximately 144 degrees, approximately 145 degrees, approximately 146 degrees, approximately 147 degrees, approximately 148 degrees, approximately 149 degrees, approximately 150 degrees, approximately 151 degrees, approximately 152 degrees, approximately 153 degrees, approximately 154 degrees, approximately 155 degrees, approximately 156 degrees, approximately 157 degrees, approximately 158 degrees, approximately 159 degrees, approximately 160 degrees, approximately 161 degrees, approximately 162 degrees, approximately 163 degrees, approximately 164 degrees, approximately 165 degrees, approximately 166 degrees, approximately 167 degrees, approximately 168 degrees, approximately 169 degrees, or approximately 170 degrees.

[0043] As described above, the light emitter may be an optical fiber and may include a source of laser light (e.g., a monochromatic light source). The system may include one or more light emitters. When multiple light emitters are used, the multiple light emitters may be connected to the same or different laser sources. In some variations, one or more light emitters may include two light emitters, namely a first light emitter and a second light emitter. The first and second light emitters may be configured within the system such that the first light emitter is positioned above the sample plate and the second light emitter is positioned below the sample plate. For example, as illustrated in FIG. 2a, a first module (200) (configured to emit light and detect scattered light at least) may include a first light emitter (202) positioned above the sample plate (204) and a second light emitter (206) positioned below the sample plate (204). Illumination by the first and second light emitters may be sequential (i.e., not simultaneous). A first light emitter (202) illuminating a sample in a sample plate (204) from the top may cause backlight scattering. A second light emitter (206) illuminating a sample in a sample plate (204) from the bottom may cause forward light scattering.

[0044] As previously mentioned, the system may include a single photodetector configured to receive light scattered at multiple scattering angles. The single photodetector may be connected to a photon counter and may have a detector beam axis parallel to the longitudinal axis of the photodetector. In some variations, the single photodetector may be an optical fiber coupled to a rotary actuator in such a way that the optical fiber rotates when the actuator rotates around the actuator's axis of rotation (e.g., the actuator's longitudinal axis). For example, referring to FIGS. 2a and 2b, a single photodetector (208) positioned on a sample plate (204) may be coupled to an actuator (210) configured to rotate around a axis of rotation (212). More specifically, a single photodetector (208) can be coupled to the actuator (210) via a clamp (214) on the swing arm (216) of the actuator (210). As shown in FIG. 2c, the actuator (210) can rotate the photodetector (208) in the direction of the arrow (218). The rotation can be between about 1.0 degree and about 280 degrees around the axis of rotation (212), including all values ​​and sub-ranges within it. In some variations, the angle of the swing arm (216) can be moved in increments of about 1.8 degrees. Such a wide angle sweep can allow a larger amount of scattered light to be detected and analyzed, thus leading to a more accurate determination of the size of particles within the sample. In one variation, rotating the photodetector and selecting illumination by a first (e.g., above the sample plate) and / or a second (e.g., below the sample plate) photoemitter may be included in the procedure for setting the scattering angle at which light scattering is detected for the sample. The illumination from the photoemitter may have an emission beam axis parallel to the longitudinal axis of the photoemitter from which the illumination is generated.

[0045] As further described below, the sample plate may include multiple cuvettes containing small volume samples for characterization / analysis. The first module may be configured to adjust the position of the emission beam axis so that the intersection (beam overlap) between the emission beam axis and the detector beam axis is maintained. Adjustment may be achieved using one or more actuators, such as stepper motors. The actuators may be adjusted manually or automatically based on instructions from one or more system processors. Considering that any deformation of the sample plate (e.g., the surface of the sample plate) may alter the optical path of the emission beam and / or the reception beam (including light scattered at a specific scattering angle), it may be useful to restore the intersection of the emission beam and the reception beam so that beam overlap occurs at the same location whenever the photodetector is moved (e.g., rotated) and / or whenever the illumination direction is changed (e.g., from top to bottom or vice versa) (this may help improve the accuracy of the acquired data).

[0046] For example, referring to FIG. 3a, it may be useful to restore the beam overlap of the emission beam (300) and the reception beam (302) so that the overlap forms a conical shape as the photodetector rotates. If the beam overlap is not restored, the emission beam (304) and the reception beam (306) may form a hyperbolic surface as shown in FIG. 3b. Defects in the swing arm and the photodetector facet may also contribute to deformation of the optical paths of the emission beam and the reception beam. The first module may include a stage configured to move one or more photoemitters in a horizontal plane in the X' and / or Y' direction so that the beam overlap can be restored. For example, referring to FIG. 3c, the first module (308) may include a stage (310) comprising a first stage actuator (310a) configured to translate a light emitter (312) (one light emitter positioned above the sample plate (314) and one light emitter positioned below the sample plate (314)) in the Y' direction, and a second stage actuator (310b) configured to translate the light emitter (312) in the X' direction.

[0047] The first module may further include a third stage actuator (e.g., a stepper motor) configured to translate a photodetector and / or photoemitter in the Z' direction in a vertical plane so that the intersection (beam overlap) of the emitted beam and the received beam occurs at the same height in a plurality of cuvettes. For example, referring to FIG. 4a, the first module (400) may include a third stage actuator (408) configured to translate the first module (400) and the associated single photodetector (402) and / or photoemitter (404) (one photoemitter positioned above the sample plate (406) and one photoemitter positioned below the sample plate (406)) in the upward and downward directions, e.g., in the Z' direction. As further detailed in FIGS. 4b and 4c, the beam overlap (410) of the emitting beam (412) and the receiving beam (414) is maintained at the same height (H) in the cuvette (416). FIG. 4b illustrates the beam overlap (410) when illuminated from the light emitter (418) above the sample plate (420), and FIG. 4c illustrates the beam overlap (410) when illuminated from the light emitter (422) below the sample plate (420). In one variation, when the cuvette is 0.7 mm high, it may be useful to maintain the intersection point at a height of 0.35 mm in each cuvette of the plurality of cuvettes.

[0048] 2nd Module (Sample Plate Positioning Module)

[0049] Some variations of the system described herein may include a second module configured to move a sample plate to a position where light scattering and / or UV / Vis measurements are to be acquired. For example, referring to FIG. 1b, the second module (106) may include a microplate drawer (110) configured to accommodate a sample plate (114) within it and to adjust the position of the sample plate (114) within the housing (102) to acquire light scattering and / or absorbance data. The second module may be configured for manual or automatic translational movement. One or more second module actuators (e.g., motors) may be used to implement the positioning of the sample plate. For example, one or more second module actuators may be used to move the sample plate so that each cuvette is properly positioned to acquire a measurement by a single photodetector and / or to move the plate so that each cuvette of the sample plate can be analyzed sequentially. When position adjustment is automatic, the second module may be configured to receive instructions from one or more processors of the system based on information entered by the user or based on data / information from, for example, system calibration, type(s) of the sampled particles, size of the cuvette, etc. A single photodetector may rotate over a sample plate holding multiple samples to detect light scattered at multiple scattering angles.

[0050] The sample plate positioned within the second module may be disposable and configured in various ways, but generally may be structured to hold one or more samples. When the plate contains multiple samples, the plate may hold 2 to 96 samples. For example, the plates are 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 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, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, or 96 samples can be held. In some variations, the sample plate can be designed for automated, high-throughput use (e.g., by utilizing a stackable design and including a barcode for tracking). The sample plate can also be configured for manual pipetting of samples or configured to be compatible with liquid handling robots that feed samples into the wells of the plate.

[0051] The sample plate may have a length ranging from about 11 cm to about 13 cm, including all values ​​and sub-ranges within it. For example, the length of the sample plate may be about 11 cm, about 11.5 cm, about 12 cm, about 12.5 cm, or about 13 cm. The width of the sample plate may have a width ranging from about 7.0 cm to about 9.0 cm, including all values ​​and sub-ranges within it. For example, the width of the sample plate may be about 7.0 cm, about 7.5 cm, about 8.0 cm, about 8.5 cm, or about 9.0 cm. In one variation, the sample plate may have a length of about 12.8 cm and a width of about 8.5 cm. Regarding material, the sample plate may be made of a polymer (e.g., a thermoplastic polymer), plastic, metal, glass, or a combination thereof. In one variation, the sample plate can be made of acrylonitrile butadiene styrene (ABS).

[0052] In some variations, the sample plate may include a plurality of cuvettes, and within the plurality of cuvettes, a sample is included to acquire light scattering and / or absorbance data. The plurality of cuvettes may be placed on a member configured as a strip (referred herein to as a “chip” or “microfluidic chip”). The sample plate may include one strip or a plurality of strips. For example, the sample plate may include one, two, three, four, five, six, seven, eight, nine, or ten strips. In some variations, it may be advantageous for the sample plate to include six strips. One or more strips may be pre-mounted on the substrate of the plate.

[0053] The strip may have a length and a width. The length of the strip may range from about 7.0 cm to about 9.0 cm, including all values ​​and sub-ranges therein. For example, the strip may have a length of about 7.0 cm, about 7.5 cm, about 8.0 cm, about 8.5 cm, or about 9.0 cm. The width of the strip may range from about 1.0 cm to about 3.0 cm, including all values ​​and sub-ranges therein. For example, the width of the strip may be about 1.0 cm, about 1.5 cm, about 2.0 cm, about 2.5 cm, or about 3.0 cm. In one variation, it may be useful for the strip to have a length of about 8.1 cm and a width of about 1.8 cm. The strip may be made of any suitable material, including polymers (e.g., thermoplastic polymers), plastics, glass, and combinations thereof, without limitation. In one variation, the strip can be made of a cyclic olefin copolymer (COC). In another variation, the strip can be made of glass. In a further variation, the strip can be formed of a material that makes the strip transparent.

[0054] Multiple wells configured to receive samples (e.g., input wells) may also be included on the strip. The strip may include any number of wells. For example, the strip may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 wells. The wells may have a conical shape to fit the tip of a pipette, but other shapes may be used. The samples input into the multiple wells may be the same or different. The sample volume may be very small, ranging from about 0.1 μl to about 2.5 μl, including all values ​​and sub-ranges within it. For example, the sample volume may be about 0.1 μl, about 0.2 μl, about 0.3 μl, about 0.4 μl, about 0.5 μl, about 0.6 μl, about 0.7 μl, about 0.8 μl, about 0.9 μl, about 1.0 μl, about 1.1 μl, about 1.2 μl, about 1.3 μl, about 1.4 μl, about 1.5 μl, about 1.6 μl, about 1.7 μl, about 1.8 μl, about 1.9 μl, about 2.0 μl, about 2.1 μl, about 2.2 μl, about 2.3 μl, about 2.4 μl, or about 2.5 μl. In one variation, the volume of at least one sample may be about 2.0 μl. In another variation, the volume of at least one sample may be about 2.0 μl or less. The sample may contain one or more types of particles. As previously described herein, one or more types of particles may be, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. The size of one or more types of particles in the sample may range from about 0.3 nm to about 3,000 nm, including all values ​​and sub-ranges therein. For example, the size of one or more particles may be about 0.3 nm, about 0.4 nm, about 0.5 nm, about 0.6 nm, about 0.7 nm, about 0.8 nm, about 0.9 nm, about 1.0 nm, approx. 1.5 nm, approx. 2.0 nm, approx. 2.5 nm, approx. 3.0 nm, approx. 3.5 nm, approx. 4.0 nm, approx. 4.5 nm, approx. 5.0 nm, approx. 5.5 nm, approx. 6.0 nm, approx. 6.5 nm, approx. 7.0 nm, approx. 7.5 nm, approx. 8.0 nm, approx. 8.5 nm, approx. 9.0 nm, approx. 9.5 nm, approx. 10 nm, approx. 15 nm, approx. 20 nm, approx. 25 nm, approx. 30 nm, approx. 35 nm, approx. 40 nm, approx. 45 nm, approx. 50 nm, approx. 55 nm, approx. 60 nm, approx. 65 nm, approx. 70 nm, approx. 75 nm, approx. 80 nm, approx. 85 nm, approx. 90 nm, approx. 95 nm, approx. 100 nm, approx. 150 nm, approx. It may be 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 1,000 nm, about 1,500 nm, about 2,000 nm, about 2,500 nm, or about 3,000 nm.

[0055] The strip may be configured so that a sample placed in a well flows through at least one channel, such as at least one microchannel, into one or more cuvettes for analysis. The cuvettes may have a path length ranging from about 0.1 mm to about 0.7 mm, including all values ​​and sub-ranges within them. For example, the path length may be about 0.1 mm, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, or about 0.7 mm. In some variations, the path length of the cuvette is about 0.1 mm. In other variations, the path length of the cuvette is about 0.7 mm.

[0056] In some variations, the sample plate may be configured as illustrated in FIG. 5a. Referring to FIG. 5a, the sample plate (500) may include a plurality of strips, such as six strips (502), on a substrate (504). A plan view of one of the six strips (502) is provided separately in FIG. 5b, which illustrates the configuration of a plurality of wells (504) placed on the strip (502). An enlarged view of the area (E) indicated by the box is provided in FIG. 5c. In FIG. 5c, one of the plurality of wells (504) is illustrated as being fluidly connected to at least one cuvette (506) through a channel (508). The cuvette (506) is illustrated in a shape such as a rounded rhombus, but other shapes may be used. For example, the cuvette may be shaped into a circular, elliptical, triangular, square, or rectangular shape. Referring to FIGS. 6a through 6d, a sample (yellow) is placed in a well (600) (Fig. 6a), and then drawn into a channel (602) by capillary force to prevent evaporation (Fig. 6b). Next, empty cuvettes (604, 606) are read (Fig. 6c). Then, a small vacuum pressure (e.g., about 20 mm Hg) from one or more pumps of the system is applied to a vent hole (608) to move the sample into the cuvette (604) and / or cuvette (606), and light scattering measurements can be obtained (Fig. 6d). The pumps may be contained within the housing of the system or located outside the housing of the system.

[0057] 3rd Module (Spectrometer Module)

[0058] In some variations, the system described herein may include a third module configured to perform UV / Vis spectroscopy. For example, referring to FIGS. 1a and FIGS. 1b, the third module (108) may be located adjacent to the second module (106) and the first module (104) within the housing (102). Thus, in this variation, the system may measure both the absorbance of the sample and light scattering at multiple angles as a function of wavelengths, for example, in the range of 230 nm to 750 nm. From the measured UV / Vis absorption spectrum, the quantity and quality of the particles may be calculated. Dynamic light scattering measurements obtained at multiple angles using a single rotating photodetector of the first module may be used for particle size determination and / or aggregation detection as described herein.

[0059] When in use, absorbance measurements can be obtained from the sample after it has been transferred to one or more cuvettes. Thus, referring again to FIGS. 6a through 6d, when the sample is transferred to cuvette (604) and / or cuvette (606), absorbance measurements can be obtained using the spectrometer of the third module. Absorbance measurements can be obtained before or after light scattering measurements (described above) are obtained. In some cases, the system can simultaneously measure UV / Vis absorption through the cuvette during the pressure-driven transfer of the sample from the channel to the cuvette. This may allow the system to monitor the filling behavior of the cuvette in addition to analyzing the spectral absorbance of the sample.

[0060] The system may include any number of modules or combinations of modules, but generally will include a first module, a second module, and a third module as described herein. For example, the system may include a first module for measuring light scattering to obtain particle size(s) and / or particle size distribution, a second module for ensuring that sample plates and cuvettes can be properly positioned when performing measurements, calibration, and / or obtaining other data, and a third module for measuring the absorbance of sample(s). The first, second, and third modules may be contained within the housing of the system, as illustrated in FIGS. 1a and 1b. Other system components, such as a vacuum pump, laser source(s), and a processor / processing unit, may also be contained within the housing. However, in some variations, one or more processor / processing units may be located outside the housing (e.g., within a computer) and may be telegraphically coupled to the system modules, for example, via a wired or wireless connection. As previously mentioned, data from scattered light at multiple (e.g., different) angles may be combined and analyzed by one or more processors to determine a single result that may be independent of the angle. In some variations, one or more processors may utilize an algorithm configured to determine particle size using data and / or signals related to one or more of sample temperature, sample viscosity, hydrodynamic diameter of the particle, intensity of scattered light, ratio of scattered light intensity to incident light intensity at a specific angle, angle of scattered light, and rate of change of scattered light intensity (e.g., variation of scattered light intensity over time). In some variations, one or more processors may include a correlator configured to analyze particle size based on data and / or intensity of various signals described herein.

[0061] Additionally, the system may include a display configured to show / present to the user various types of information / data, such as system status, warnings, sample plate information, sample and / or particle information, light scattering measurements, particle size, particle size distribution, absorbance measurements, other data related to any of the foregoing, graphical representations of measurements / data, etc. The display may be placed on a part of the housing or may include a monitor coupled to the system. The display may also include a user interface or function as a user interface, thereby enabling the user to control one or more modules and / or components of the system. In this example, the user interface may include a touch screen. In other variations, the user interface may include buttons, a keyboard, or a keypad that the user can press to interact with and control various components of the system.

[0062] One or more processors may be configured to control the operation of system modules, other system components, and their interactions. Accordingly, one or more processors may be configured to execute instructions for performing one or more of determining the size and quantifying particles or other materials within a sample. For example, one or more processors of the system may be configured to determine the size of one or more types of particles within a sample based on data from scattered light. As previously mentioned, the size of one or more types of particles within a sample may be in the range of about 0.3 nm to about 3,000 nm, including all values ​​and sub-ranges therein. One or more processors may be configured to receive multiple signals from a single photodetector, where each of the multiple signals may represent scattered light at a specific angle. Then, as further described below, data from scattered light at multiple (e.g., different) angles may be combined and analyzed by one or more processors to determine a single result that may be independent of the angle.

[0063] method

[0064] The present specification also describes a method for characterizing particles. Exemplary particles include, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. Generally, the method may include the steps of illuminating a plurality of samples with light from one or more light emitters—wherein each of the plurality of samples contains one or more types of particles—; rotating a photodetector over a plurality of scattering angles around a rotation axis; detecting light scattered by particles over a plurality of scattering angles; and obtaining light scattering measurements for each of the plurality of scattering angles. The rotation of the photodetector may occur over the plurality of samples (i.e., over a sample plate). As mentioned above, data from the scattered light at a plurality of scattering angles may then be combined and analyzed by one or more processors to determine a single result that may be independent of the angle.

[0065] One or more light emitters may include a laser light source. It is understood that suitable alternative light sources may also be used. Some variations of the method may utilize a first light emitter and a second light emitter. In such variations, the first light emitter may be placed above a plurality of samples, and the second light emitter may be placed below a plurality of samples. When two light emitters are used, e.g., a first light emitter above a sample plate and a second light emitter below a sample plate, illumination by each light emitter is generally sequential. In other words, the first light emitter may illuminate each of the plurality of samples, followed by illumination by the second light emitter, or vice versa.

[0066] Rotation can be achieved by using an actuator that rotates a photodetector when the actuator is rotated around its axis of rotation (e.g., its longitudinal axis). When the photodetector is rotated to acquire scattered light at multiple angles, the angle of rotation may be in the range of about 1 degree to about 280 degrees, including all values ​​and sub-ranges within the axis of rotation of the actuator. For example, a single photodetector rotates about 1.0°, 5.0°, 10°, 15°, 20°, 25°, 30°, 35°, 40°, 45°, 50°, 55°, 60°, 65°, 70°, 75°, 80°, 85°, 90°, 95°, 100°, 105°, 110°, 115°, 120°, 125°, 130°, 135°, 140°, 145°, 150°, 155°, 160°, 165°, 170°, 175°, and approximately around the axis of rotation. It can rotate 180 degrees, approximately 185 degrees, approximately 190 degrees, approximately 195 degrees, approximately 200 degrees, approximately 205 degrees, approximately 210 degrees, approximately 215 degrees, approximately 220 degrees, approximately 225 degrees, approximately 250 degrees, approximately 255 degrees, approximately 260 degrees, approximately 265 degrees, approximately 270 degrees, approximately 275 degrees, or approximately 280 degrees. In some variations, a single photodetector can rotate 280 degrees around the axis of rotation or more than 280 degrees around the axis of rotation. In other variations, rotation of the actuator between approximately 30 degrees and approximately 180 degrees can also rotate the single photodetector by a corresponding angle. This wide range of angle sweeps can allow a larger amount of scattered light to be detected and analyzed, which can thus lead to more accurate determination of the size of particles within the sample.

[0067] Both forward scattered light and backscattered light can be detected by a single rotating light detector. Forward light scattering can be detected at multiple scattering angles in the range of about 25 degrees to about 45 degrees, including all values ​​and sub-ranges therein. For example, the forward light scattering angles of the multiple scattering angles can be about 25 degrees, about 26 degrees, about 27 degrees, about 28 degrees, about 29 degrees, about 30 degrees, about 31 degrees, about 32 degrees, about 33 degrees, about 34 degrees, about 35 degrees, about 36 degrees, about 37 degrees, about 38 degrees, about 39 degrees, about 40 degrees, about 41 degrees, about 42 degrees, about 43 degrees, about 44 degrees, or about 45 degrees.

[0068] Backlight scattering can be detected at multiple scattering angles ranging from about 105 degrees to about 170 degrees, including all values ​​and sub-ranges within it. For example, the backlight scattering angles of multiple scattering angles are approximately 105°, approximately 106°, approximately 107°, approximately 108°, approximately 109°, approximately 110°, approximately 111°, approximately 112°, approximately 113°, approximately 114°, approximately 115°, approximately 116°, approximately 117°, approximately 118°, approximately 119°, approximately 120°, approximately 121°, approximately 122°, approximately 123°, approximately 124°, approximately 125°, approximately 126°, approximately 127°, approximately 128°, approximately 129°, approximately 130°, approximately 131°, approximately 132°, approximately 133°, approximately 134°, approximately 135°, approximately 136°, approximately 137°, approximately It may be 138 degrees, approximately 139 degrees, approximately 140 degrees, approximately 141 degrees, approximately 142 degrees, approximately 143 degrees, approximately 144 degrees, approximately 145 degrees, approximately 146 degrees, approximately 147 degrees, approximately 148 degrees, approximately 149 degrees, approximately 150 degrees, approximately 151 degrees, approximately 152 degrees, approximately 153 degrees, approximately 154 degrees, approximately 155 degrees, approximately 156 degrees, approximately 157 degrees, approximately 158 degrees, approximately 159 degrees, approximately 160 degrees, approximately 161 degrees, approximately 162 degrees, approximately 163 degrees, approximately 164 degrees, approximately 165 degrees, approximately 166 degrees, approximately 167 degrees, approximately 168 degrees, approximately 169 degrees, or approximately 170 degrees.

[0069] In some variations, the method may further include the step of adjusting the position of the emission beam axis so that the intersection point (beam overlap) between the emission beam axis and the detector axis is restored at each cuvette when light scattering is measured at multiple scattering angles and / or when the illumination direction is changed (e.g., from the top to the bottom of the plate or vice versa). In other words, light scattering measured at a first scattering angle has a first beam overlap, and light scattering measured at a second scattering angle has a second beam overlap that can be restored to be identical or substantially identical to the first beam overlap, as illustrated in FIG. 3a. Light scattering measured at subsequent scattering angles (e.g., third, fourth, fifth, sixth, seventh, and eighth scattering angles) can also be restored so that the beam overlap is identical or substantially identical, and accordingly, as illustrated by the comparison between FIG. 3a (cone) and FIG. 3b (hyperbolic surface), the beam overlap forms a cone rather than a hyperbolic shape. The position of the emission beam axis can be adjusted manually or automatically according to instructions from one or more processors. Restoration of beam overlap may be necessary to account for factors such as defects in the swing arm coupled to the rotary actuator or photodetector face and / or deformation of the strip surface that may deform the optical paths of the emission and reception beams, including scattered light.

[0070] The adjustment of the emission beam axis to restore beam overlap (e.g., automatic adjustment) may be based on an algorithm (e.g., processing / method step) executed by one or more processors, which includes identifying a maximum intensity measurement. The algorithm may include one or more preliminary positioning steps. For example, as illustrated in FIGS. 7a through 7d, one or more preliminary positioning steps may include moving one or more photo-emitters to theoretical positions on a hyperbolic surface, e.g., the smallest circle, during calibration of the system ( FIG. 7a). Next, the algorithm may include one or more fine positioning steps. In some variations, fine positioning may include the step of moving the photo-emitters (2D sweep) to find a desired overlap (e.g., maximum intensity), as illustrated in FIG. 7b, and generating a 2D plot of intensity. Referring to FIGS. 7a and 7b, the intensity may be constant when moving along line A, which is the bisector between the photo-emitter and the photo-detector. Additionally or alternatively, micro-localization may further include a step of scanning the intensity along line A to find the maximum intensity. For example, FIG. 7c illustrates the measured intensity along line A. Given that the intensity along line A is known to be Gaussian, it is sufficient to measure a limited number of points (e.g., 4, 5, 6, or 7 points). For each point (indicated by 5 black dots), as shown in FIG. 7d, the point can be fitted to a Gaussian curve and / or its location can be refined, the maximum value of the curve can be identified, and the intensity at the maximum value can be measured.

[0071] The method may also include the step of maintaining beam overlap at the same height in each cuvette while light scattering measurements are acquired from a plurality of cuvettes. In some variations, the same height may be maintained using an actuator, for example, a third-stage actuator configured to translate a photodetector and / or photoemitter in the Z' direction in the vertical plane so that the intersection of the emitted beam and the received beam (beam overlap) occurs at the same height in a plurality of cuvettes as described above. The steps involved in maintaining beam overlap may be as illustrated in FIGS. 4a through 4c. Referring to FIG. 4a, the first module (400) may include a third stage actuator (408) configured to translate the first module (400) and the associated single photodetector (402) and / or photoemitter (404) (one photoemitter positioned above the sample plate (406) and one photoemitter positioned below the sample plate (406)) in the upward and downward directions, e.g., in the Z' direction. As further detailed in FIG. 4b and FIG. 4c, the beam overlap (410) of the emitting beam (412) and the receiving beam (414) is maintained at the same height (H) in the cuvette (416). FIG. 4b illustrates beam overlap (410) when illuminated from a light emitter (418) above the sample plate (420), and FIG. 4c illustrates beam overlap (410) when illuminated from a light emitter (422) below the sample plate (420). In one variation, when the cuvette is 0.7 mm high, it may be useful to maintain the intersection (beam overlap) at a height of 0.35 mm in each cuvette of the plurality of cuvettes. The beam overlap can be adjusted manually or automatically according to instructions from one or more processors.

[0072] Additionally, the method may include the step of acquiring absorbance data (e.g., UV / Vis data) from a plurality of samples. For example, the plurality of samples may be illuminated with light having a wavelength in the range of about 190 nm to about 900 nm, and the absorbance may be measured. The absorbance data may be acquired using a module of the system, such as a second module as previously described. The absorbance data may be acquired before or after the light scattering data is acquired.

[0073] Characterization of particles can be achieved for small sample volumes ranging from about 0.5 μl to about 2.5 μl, including all values ​​and sub-ranges within them. For example, the sample volume may be about 0.1 μl, about 0.2 μl, about 0.3 μl, about 0.4 μl, about 0.5 μl, about 0.6 μl, about 0.7 μl, about 0.8 μl, about 0.9 μl, about 1.0 μl, about 1.1 μl, about 1.2 μl, about 1.3 μl, about 1.4 μl, about 1.5 μl, about 1.6 μl, about 1.7 μl, about 1.8 μl, about 1.9 μl, about 2.0 μl, about 2.1 μl, about 2.2 μl, about 2.3 μl, about 2.4 μl, or about 2.5 μl. In one variation, the volume of multiple samples is about 2 μl. In another variation, the volume of multiple samples is about 2 μl or less.

[0074] Determining the size of one or more types of particles within a sample can be achieved by one or more processors of a system configured to analyze data from scattered light. The size of one or more types of particles within a sample may be in the range of about 0.3 nm to about 3,000 nm, including all values ​​and sub-ranges therein. For example, the size of one or more particles is approximately 0.3 nm, approximately 0.4 nm, approximately 0.5 nm, approximately 0.6 nm, approximately 0.7 nm, approximately 0.8 nm, approximately 0.9 nm, approximately 1.0 nm, approximately 1.5 nm, approximately 2.0 nm, approximately 2.5 nm, approximately 3.0 nm, approximately 3.5 nm, approximately 4.0 nm, approximately 4.5 nm, approximately 5.0 nm, approximately 5.5 nm, approximately 6.0 nm, approximately 6.5 nm, approximately 7.0 nm, approximately 7.5 nm, approximately 8.0 nm, approximately 8.5 nm, approximately 9.0 nm, approximately 9.5 nm, approximately 10 nm, approximately 15 nm, approximately 20 nm, approximately 25 nm, approximately 30 nm, approximately 35 nm, approximately 40 nm, approximately 45 nm, approximately 50 nm, approximately 55 nm, approximately It may be 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 1,000 nm, about 1,500 nm, about 2,000 nm, about 2,500 nm, or about 3,000 nm. As previously mentioned in this specification, particles may include, without limitation, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof.

[0075] In some variations, data from scattered light at multiple (e.g., different) angles may be combined and analyzed by one or more processors to determine a single result that may be independent of the angle. For example, one or more processors may utilize an algorithm configured to determine particle size using data and / or signals related to one or more of sample temperature, sample viscosity, hydrodynamic diameter of the particle, intensity of scattered light, ratio of scattered light intensity to incident light intensity at a specific angle, angle of scattered light, and rate of change of scattered light intensity (e.g., variation of scattered light intensity over time). In some variations, one or more processors may include a correlator configured to analyze particle size based on the data and / or intensity of the various signals described above.

[0076] In some variations, when a signal is acquired from scattered light detected at multiple scattering angles by a single photodetector, the data from the scattered light at multiple (e.g., different) angles is then combined and analyzed by one or more processors to determine (e.g., use to calculate) a single result (e.g., a single solution) characterizing particles within a sample (e.g., particle size and / or particle size distribution). The single solution may be angle-independent. In one variation, determining the single solution involves simultaneously satisfying all desired absolute correlation functions, and the single solution is expressed by the following equation:

[0077]

[0078] R(q) is the scatter intensity to Rayleigh ratio for each angle (Rayleigh ratio at angle q), and R(q) is multiplied by the square root of the correlation function. The correlation function can be transformed to include adjustments for different scattering angles, temperatures, and viscosities (e.g., by rescaling the x-axis).

[0079] In some variations, one or more processors may use absolute correlation data obtained from light scattered at multiple angles to generate a graphical representation of the absolute correlation function, such as a 2D representation or a model. For a single particle, the 2D graphical representation may be a plane having a slope in the x-direction and the q²-direction. Thus, for a mixture of particles, the 2D graphical representation may include the sum of these planes (e.g., the planes are presented collectively). In one variation, the 2D representation may help determine the size of one or more types of particles within each sample based collectively on the shape of the absolute correlation function of multiple scattering angles, as illustrated in FIGS. 8a through 8c. For example, when the sample contains relatively small particles (e.g., 10 nm), the shape of the absolute correlation function may be as illustrated in FIG. 8a, where all planes are parallel, there is rapid decay in the x-direction, and no decay in the q²-direction. When the sample contains relatively larger particles (e.g., 100 nm), the shape of the absolute correlation function may be as shown in FIG. 8b, where all planes are also parallel, but there is slow decay in the x-direction and fast decay in the q²-direction. When the sample contains a mixture of particles (e.g., 10 nm and 100 nm), the shape of the absolute correlation function may be as shown in FIG. 8c, where the planes are not parallel but spread out in a fan shape in the x-direction.

[0080] Although the foregoing variations have been described in some detail through examples and illustrations for the purposes of clarity and understanding, it will be apparent that specific changes and modifications may be made, and such changes and modifications are intended to fall within the scope of the appended claims. Additionally, it should be understood that the components and characteristics of the systems and devices described herein may be used in any combination. Descriptions of specific elements or characteristics in relation to specific drawings are not intended to be limiting, nor should they be interpreted as implying that the element cannot be used in combination with any of the other elements described. For all variations described herein, the steps of the method may not be performed sequentially. Some steps are optional, and accordingly, not all steps of the method may be performed.

Claims

Claim 1 A system for characterizing particles comprises: one or more processors; a plurality of samples—each of the plurality of samples comprises one or more types of particles—; and a first module configured to measure light scattered by each sample, wherein the first module comprises, One or more light emitters; An actuator having a rotation axis; and A system for characterizing particles comprising a single photodetector configured to rotate around the above-mentioned rotation axis and receive light scattered by each sample at a plurality of scattering angles. Claim 2 A system for characterizing particles according to claim 1, further comprising a sample plate configured to hold the plurality of samples. Claim 3 A system for characterizing particles according to paragraph 2, wherein the sample plate comprises a plurality of cuvettes. Claim 4 A system for characterizing particles according to paragraph 3, wherein the first module comprises one or more stage actuators configured to adjust the position of the emission beam axis so that the intersection [beam overlap] between the emission beam axis and the detector beam axis is restored within each sample when the photodetector receives light scattered at each angle of the plurality of scattering angles. Claim 5 A system for characterizing particles according to claim 4, wherein the one or more stage actuators are configured to automatically adjust the position of the emission beam axis. Claim 6 A system for characterizing particles according to claim 4, wherein the one or more stage actuators are also configured to adjust the position of the first module so that the intersection point (beam overlap) between the emission beam axis and the detector beam axis is maintained at the same height in the plurality of cuvettes. Claim 7 A system for characterizing particles according to paragraph 2, further comprising a second module configured to automatically adjust the position of the sample plate according to instructions from one or more processors. Claim 8 A system for characterizing particles according to claim 1, wherein the single photodetector comprises an optical fiber. Claim 9 A system for characterizing particles according to claim 1, wherein the single photodetector is configured to rotate about 1.0 degree to about 280 degrees around the rotation axis. Claim 10 A system for characterizing particles according to claim 1, wherein the single photodetector is configured to receive forward light scattering from the plurality of scattering angles. Claim 11 A system for characterizing particles according to claim 10, wherein the plurality of scattering angles are in the range of about 25 degrees to about 45 degrees. Claim 12 A system for characterizing particles according to claim 1, wherein the single photodetector is configured to receive backward light scattering from the plurality of scattering angles. Claim 13 A system for characterizing particles according to claim 12, wherein the plurality of scattering angles are in the range of about 105 degrees to about 170 degrees. Claim 14 A system for characterizing particles according to claim 1, further comprising a third module configured to illuminate the plurality of samples with light having a wavelength in the range of about 190 nm to about 900 nm and to measure the absorbance of the plurality of samples. Claim 15 A system for characterizing particles according to claim 1, wherein one or more light emitters include a laser light source. Claim 16 A system for characterizing particles according to claim 1, wherein the one or more light emitters include a first light emitter and a second light emitter. Claim 17 A system for characterizing particles according to claim 16, wherein the first light emitter is positioned above the sample plate and the second light emitter is positioned below the sample plate. Claim 18 A system for characterizing particles according to claim 1, wherein one or more types of particles include nanoparticles, DNA, RNA, viruses, proteins, polymers, or small molecules. Claim 19 A system for characterizing particles according to claim 1, wherein each of the plurality of samples is about 2 μl or less. Claim 20 A system for characterizing particles according to paragraph 3, wherein the sample plate comprises a plurality of members, and each member of the plurality of members is configured as a strip having a length and a width. Claim 21 A system for characterizing particles according to claim 20, wherein the plurality of members comprises six strips. Claim 22 A system for characterizing particles according to claim 20, wherein the strip comprises a plurality of wells configured to accommodate the plurality of samples. Claim 23 A system for characterizing particles according to claim 22, wherein each well of the plurality of wells is coupled to a corresponding cuvette among the plurality of cuvettes by a channel. Claim 24 A system for characterizing particles according to claim 3, wherein each cuvette of the plurality of cuvettes has a path length in the range of about 0.1 mm to about 0.7 mm. Claim 25 A system for characterizing particles according to claim 24, wherein the path length is approximately 0.1 mm. Claim 26 A system for characterizing particles according to claim 24, wherein the path length is approximately 0.7 mm. Claim 27 A system for characterizing particles according to claim 1, wherein the one or more processors are configured to determine the size of the one or more types of particles within the sample. Claim 28 A system for characterizing particles according to claim 27, wherein the size of one or more types of particles in the sample is in the range of about 0.3 nm to about 3,000 nm. Claim 29 A system comprising: a first module comprising a photodetector configured to rotate over a sample plate containing a plurality of samples and to detect scattered light from each sample at a plurality of scattering angles due to the rotation; and a second module configured to measure the absorbance of each of the plurality of samples, wherein each of the plurality of samples contains one or more types of particles. Claim 30 A system according to claim 29, further comprising one or more processors configured to determine particle size or particle size distribution from scattered light detected at the plurality of scattering angles. Claim 31 A method for characterizing particles, comprising the steps of: illuminating a plurality of samples with light from one or more light emitters, wherein each of the plurality of samples comprises one or more types of particles; rotating a light detector over a plurality of scattering angles around a rotation axis; detecting light scattered by the particles over the plurality of scattering angles; and obtaining light scattering measurements for each of the plurality of scattering angles. Claim 32 A method for characterizing a particle according to claim 31, further comprising the step of adjusting the position of the emission beam axis so that the intersection point (beam overlap) between the emission beam axis and the detector axis is restored within each sample when the photodetector receives light scattered at each of the plurality of scattering angles. Claim 33 A method for characterizing a particle according to claim 32, further comprising the step of adjusting the position of the intersection point (beam overlap) between the emission beam axis and the detector beam axis so that the beam overlap is maintained at the same height in the plurality of cuvettes. Claim 34 A method for characterizing particles according to claim 32, further comprising the step of automatically adjusting the position of the emission beam axis. Claim 35 A method for characterizing particles according to claim 31, further comprising the step of automatically adjusting the position of a sample plate according to instructions from one or more processors. Claim 36 A method for characterizing a particle according to paragraph 33, wherein the above instruction comprises one or more preliminary positioning steps. Claim 37 A method for characterizing a particle according to claim 36, wherein the one or more preliminary positioning steps include the step of moving the one or more light emitters to the smallest circle on a hyperboloid. Claim 38 A method for characterizing particles according to claim 36, further comprising one or more micro-positioning steps. Claim 39 A method for characterizing a particle according to claim 38, wherein the one or more micro-positioning steps include the step of determining the maximum intensity of the light scattered by each sample at the plurality of scattering angles. Claim 40 A method for characterizing particles according to claim 31, wherein the light detector comprises an optical fiber. Claim 41 A method for characterizing particles according to claim 31, wherein the photodetector is configured to rotate about 1 degree to about 280 degrees around the rotation axis. Claim 42 A method for characterizing a particle according to claim 31, wherein the photodetector detects forward light scattering from the plurality of scattering angles. Claim 43 A method for characterizing particles according to claim 42, wherein the plurality of scattering angles are in the range of about 25 degrees to about 45 degrees. Claim 44 A method for characterizing a particle according to claim 31, wherein the photodetector detects backlight scattering from the plurality of scattering angles. Claim 45 A method for characterizing particles according to claim 44, wherein the plurality of scattering angles are in the range of about 105 degrees to about 170 degrees. Claim 46 A method for characterizing particles in paragraph 31, wherein illumination is achieved using a laser light source. Claim 47 A method for characterizing a particle according to claim 31, wherein the one or more light emitters include a first light emitter and a second light emitter. Claim 48 A method for characterizing particles according to claim 47, wherein the first light emitter is placed above the plurality of samples and the second light emitter is placed below the plurality of samples. Claim 49 A method for characterizing particles according to claim 31, wherein one or more types of particles include nanoparticles, DNA, RNA, viruses, proteins, polymers, or small molecules. Claim 50 A method for characterizing particles according to claim 31, wherein each of the plurality of samples is about 2 μl or less. Claim 51 A method for characterizing particles according to claim 31, further comprising the step of determining the size of one or more types of particles in the sample based on the scattered light at the plurality of scattering angles using one or more processors. Claim 52 A method for characterizing particles according to claim 51, wherein the size of one or more types of particles in the sample is in the range of about 0.3 nm to about 3,000 nm. Claim 53 A method for characterizing particles according to claim 51, wherein one or more processors determine the size of one or more types of particles in each sample based on the shape of the absolute correlation function of the plurality of scattering angles. Claim 54 A method for characterizing particles according to claim 51, wherein the one or more processors determine the size of the one or more types of particles in each sample based on a single solution derived from light scattering measurements obtained for the plurality of scattering angles.

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