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

Abstract

Described herein are systems and methods of obtaining 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 systems and methods can characterize particles in the samples by obtaining light scattering data from a plurality of angles by a single rotating photodetector. The data obtained in this way can allow for a greater dynamic range of particle sizing than when using a single angle.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 618,653, filed January 8, 2024, which is hereby incorporated in its entirety by reference. Technical Field

[0003] This application relates to systems and methods for characterizing particles (e.g., nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules). Characterization may include obtaining particle-related light scattering and / or UV / Vis data. When obtaining light scattering data, a rotating detector capable of measuring scattered light at multiple angles can be used to detect the light scattering data. Background Technology

[0004] Dynamic light scattering is a commonly known method for analyzing particles, where measurements of scattered light over time are used to determine the particle size or size distribution. Particle properties can be inferred from the temporal variations in the scattered light. For example, autocorrelation can be performed on a time series of the scattered light intensity, and then the autocorrelation function can be fitted (e.g., cumulant, CONTIN, NNLS / non-negative least squares) to determine particle properties. In other instances, a Fourier transform can be used to determine the power spectrum of the scattered light, and a similar fit to the power spectrum can be performed to determine particle properties.

[0005] Typically, 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 very uniformly in all directions. If the particles are large, more light can be scattered in the forward direction than in the backward direction. Therefore, when analyzing a mixture of particle sizes (e.g., some small and some large), the light scattering results may be distorted when measured at a single angle, or some particles may not be visible at all because each particle size produces a unique scattering pattern. In other words, the particle-related results may vary depending on which angle is used to detect the scattered light.

[0006] Therefore, methods have been developed to detect light scattered at more than one angle to obtain dynamic light scattering measurements of a sample. Some conventional systems achieve multi-angle measurements by positioning multiple photodetectors or multiple light emitters around a single sample of interest. However, these systems have several drawbacks, including, for example, a limited range of detection angles for scattered light and / or time-consuming result processing because information must be matched between multiple photodetectors and multiple light emitters.

[0007] Therefore, there is a need for new and useful systems for detecting scattered light at multiple angles. Methods for determining particle size and / or particle size distribution in a sample would also be useful. Summary of the Invention

[0008] This document describes systems and methods for obtaining dynamic light scattering data from particle samples, and in some instances, for measuring the UV / Vis absorption spectra of particle samples (e.g., microliter-sized samples). These systems and methods characterize particles in a sample by acquiring light scattering data from multiple angles. Data obtained in this way allows for particle sizing with a greater dynamic range than when using a single angle. Additionally, quantitative data about the number of particles of each size in the sample can be obtained. The systems and methods can combine absolute correlation functions at each angle to give results independent of a single angle and can help measure previously hidden particle size groups (e.g., due to weak scattering) and / or generate a more complete size distribution. In some variations, systems incorporating a single rotating photodetector (e.g., a single rotating optical fiber) can be used to obtain dynamic light scattering measurements. The single photodetector can rotate on a sample plate holding multiple samples to detect scattered light at multiple angles for particle characterization.

[0009] A system for characterizing particles may include one or more processors, multiple samples, and a first module configured to measure light scattered by each of the multiple samples. Each of the multiple samples may contain one or more types of particles. The samples may contain any type of particles that scatter light when irradiated. Exemplary types of particles may include, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof.

[0010] 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 multiple scattering angles. The one or more light emitters may include a laser source. In some variations, 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 positioned above the sample plate, and the second light emitter may be positioned below the sample plate. It should be understood that the light emitters can be configured in various other ways.

[0011] Multiple samples can be held on a sample plate of the system. More specifically, multiple samples can 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 particles. In other variations, each of the multiple samples on the sample plate may contain different types of particles. In yet another variation, each of the multiple samples may contain a mixture of particles. In still another variation, some samples on the sample plate may contain the same particles or a mixture of the same particles, while other samples on the plate may contain different types of particles or mixtures of different particles.

[0012] The first module can also be configured to adjust the position of the emitted beam axis such that, during each rotation of the photodetector and / or when the illumination direction changes (e.g., illumination from the top of the sample plate changes to illumination from below the sample plate, and vice versa), the intersection point (beam overlap) between the emitted beam axis and the detector beam axis is restored. The first module can be configured to allow automatic or manual adjustment of the position of the emitted beam axis. Additionally, the first module can be configured to include mechanisms for maintaining beam overlap at the same height in 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.

[0013] Instead of multiple photodetectors, the system described herein typically comprises a single photodetector. The single photodetector can be configured to rotate about 1.0 degree to about 280 degrees (inclusive of all values ​​and subranges) about the rotation axis of a rotating actuator. The single photodetector may include an optical fiber.

[0014] A single photodetector can be configured to receive forward light scattering from multiple scattering angles within a range of approximately 25 degrees to approximately 45 degrees (inclusive of all values ​​and subranges therein). Regarding backscattering, a single photodetector can be configured to receive backscattering from multiple scattering angles within a range of approximately 105 degrees to approximately 170 degrees (inclusive of all values ​​and subranges therein).

[0015] Some variations of the system described herein can also be configured to acquire UV / Vis data from multiple samples. In these variations, the system may include a third module configured to irradiate multiple samples with light having wavelengths in the range of about 190 nm to about 900 nm and to measure the absorbance of the multiple samples. The particles in these samples may be particles mentioned above, such as nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules.

[0016] The volume of samples analyzed by the system can be very small, ranging from about 0.5 μl to about 2.5 μl (inclusive of all values ​​and subranges). 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.

[0017] The sample plate can be of different sizes. For example, the sample plate can have a length ranging from about 11 cm to about 13 cm (inclusive of all values ​​and subranges therein), and a width ranging from about 7.0 cm to about 9.0 cm (inclusive of all values ​​and subranges therein). In one example, the sample plate can have a length of about 12.8 cm and a width of about 8.5 cm.

[0018] The sample plate can also be configured in various ways. In some variations, the sample plate may include multiple components, each of which can be configured as a strip (referred to herein as a "chip" or "microfluidic chip") having a length and width. The length of the strip can range from about 7.0 cm to about 9.0 cm (inclusive of all values ​​and subranges therein), and the width of the strip can range from about 1.0 cm to about 3.0 cm (inclusive of all values ​​and subranges therein). For example, a strip with a length of about 8.1 cm and a width of about 1.8 cm may be useful. The sample plate may contain any suitable number of strips, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In one variation, the sample plate may contain six strips.

[0019] Each strip may include multiple wells configured to receive a sample. Each of the multiple wells may be coupled to a corresponding cuvette (from multiple cuvettes) via a channel (e.g., a microchannel). The cuvettes may have a path length in the range of about 0.1 mm to about 0.7 mm (inclusive of all values ​​and subranges therein). 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.

[0020] One or more processors of the system can be configured to determine the size of one or more types of particles in a sample based on data from the scattered light. The size of one or more types of particles in the sample can range from about 0.3 nm to about 3,000 nm (inclusive of all values ​​and subranges therein). For example, the size of one or more particles can be 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 60 nm, approximately 65 nm, approximately 70 nm, approximately 75 nm, approximately 80 nm, approximately 85 nm, approximately 90 nm, etc. nm, approximately 95 nm, approximately 100 nm, approximately 150 nm, approximately 200 nm, approximately 250 nm, approximately 300 nm, approximately 350 nm, approximately 400 nm, approximately 450 nm, approximately 500 nm, approximately 550 nm, approximately 600 nm, approximately 650 nm, approximately 700 nm, approximately 750 nm, approximately 800 nm, approximately 850 nm, approximately 900 nm, approximately 1,000 nm, approximately 1,500 nm, approximately 2,000 nm, approximately 2,500 nm, or approximately 3,000 nm. One or more processors can be configured to receive multiple signals from a single photodetector, each of which can represent scattered light at a specific angle. The data from the light scattered at multiple (e.g., different) angles can then be combined and analyzed by one or more processors to determine a single result that is angle-independent. In some variations, one or more processors may employ algorithms configured to determine particle size using data and / or signals correlated with one or more of the following: sample temperature, sample viscosity, hydrodynamic diameter of the particles, scattered light intensity, the ratio of scattered light intensity to incident light intensity at a specific angle, the scattered light angle, and the rate of change of scattered light intensity (e.g., fluctuations in scattered light intensity over time). One or more processors may include correlators configured to analyze particle size based on data and / or intensity of the various signals described above.

[0021] This document also describes methods for characterizing particles. Exemplary particles include, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. Generally, the method may include: irradiating multiple samples with light from one or more light emitters, each of the multiple samples containing one or more types of particles; rotating a photodetector about a rotation axis within multiple scattering angles; detecting the light scattered by the particles within the multiple scattering angles; and obtaining light scattering measurements for each of the multiple scattering angles. As mentioned above, the data from the light scattered at the multiple scattering angles can then be combined and analyzed by one or more processors to determine individual results that are independent of the angle.

[0022] One or more light emitters may include a laser source. It should be 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 these variations, the first light emitter may be positioned above multiple samples, and the second light emitter may be positioned below multiple samples.

[0023] In some variations, the method may further include adjusting the position of the emission beam axis such that the intersection point between the emission beam axis and the detector beam axis is maintained at the same height in the multiple cuvettes contained in the sample plate. The position of the emission beam axis can be adjusted manually or automatically according to instructions from one or more processors. Similarly, the position of the sample plate can be adjusted manually or automatically.

[0024] Automatic adjustment of the emitted beam axis can be based on instructions from one or more processors that include one or more preliminary positioning steps. For example, one or more preliminary positioning steps may include 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 multiple scattering angles.

[0025] When the photodetector rotates to obtain scattered light at multiple angles, the rotation angle can be in the range of approximately 1 degree to approximately 280 degrees (inclusive) around the rotation axis of the actuator. Forward scattering can be detected at multiple scattering angles in the range of approximately 25 degrees to approximately 45 degrees (inclusive) and backward scattering can be detected at multiple scattering angles in the range of approximately 105 degrees to approximately 170 degrees (inclusive)

[0026] Additionally, the method may include obtaining UV / Vis data from multiple samples. For example, multiple samples can be irradiated with light having wavelengths in the range of approximately 190 nm to approximately 900 nm, and the absorbance can be measured.

[0027] Particle characterization can be performed on small sample volumes ranging from about 0.5 μl to about 2.5 μl (inclusive of all values ​​and subranges therein). In one variant, multiple samples have a volume of about 2 μl. In another variant, multiple samples have a volume of about 2 μl or less.

[0028] Determining the size of one or more types of particles in a sample can be accomplished by one or more processors configured to analyze data from scattered light. The size of the one or more types of particles in the sample can range from about 0.3 nm to about 3,000 nm (inclusive of all values ​​and subranges therein), as described above. Once a signal is obtained from scattered light detected by a single photodetector at multiple scattering angles, the data from light scattered at multiple (e.g., different) angles can be combined and analyzed by one or more processors to determine a single result (e.g., a single solution). The single result can be angle-independent. In some variations, one or more processors can determine the size of one or more types of particles in each sample based on the shape of an absolute correlation function of multiple scattering angles. Attached Figure Description

[0029] This patent or application document contains at least one color drawing. Upon request and payment of the necessary fees, the Patent Office will provide a copy of the publication of this patent or application with one or more color drawings.

[0030] Figure 1A A front view of the housing of an exemplary system for measuring light scattering at multiple angles.

[0031] Figure 1B Depicting Figure 1A The modules inside the shell shown in the image.

[0032] Figure 2A A three-dimensional diagram depicting an exemplary first module (dynamic light scattering module) of a system for measuring light scattering at multiple angles.

[0033] Figure 2B and 2C An exemplary actuator is depicted, configured as a rotating photodetector to enable the detection of scattered light at multiple scattering angles.

[0034] Figure 3A and 3B An example depicting beam overlap between the emitted and received beams as the photodetector rotates. Figure 3A In the process, the beam overlap is restored. Figure 3B In the middle, the beam overlap has not yet been restored.

[0035] Figure 3CAn exemplary first module is shown, comprising a stage configured to move one or more light emitters in a horizontal plane in the X' and / or Y' directions, such that recoverable beam overlap is achieved.

[0036] Figure 4A An exemplary first module comprising an actuator configured to translate the entire first module in a vertical direction is depicted, such that beam overlap can occur at the same height in one or more cuvettes.

[0037] Figure 4B and 4C Showing when from the top of the sample plate ( Figure 4B ) and from the bottom of the sample plate ( Figure 4C When irradiating a cuvette, light beams at the same height within the cuvette overlap.

[0038] Figure 5A Describe an exemplary sample panel.

[0039] Figure 5B The exhibition is provided by Figure 5A One of the strips on the sample plate.

[0040] Figure 5C for Figure 5B An enlarged view of the structure of the strips shown in the image.

[0041] Figures 6A to 6D Describe the exemplary sample characterization cycle of the system.

[0042] Figures 7A to 7D An exemplary algorithm for restoring beam overlap is shown.

[0043] Figures 8A to 8C An exemplary 2D representation of an absolute correlation function, the shape of which helps determine the size of one or more types of particles in a sample. Detailed Implementation

[0044] This document describes systems and methods for obtaining dynamic light scattering data from particle samples, and in some cases, measuring the UV / Vis absorption spectra of the particle samples. The systems and methods characterize particles in a sample by acquiring light scattering data from multiple angles using a single photodetector that rotates due to the rotation of an actuator to which it is coupled. Data obtained in this way allows for particle size determination with a greater dynamic range than when using a single angle. The systems and methods can combine absolute correlation functions at each angle to give results independent of a single angle. Furthermore, the systems and methods can be designed for manual, automatic, or both types of workflows (e.g., switching between manual and automatic, and vice versa).

[0045] system

[0046] The systems described herein may comprise various modules that can individually or collectively aid in the characterization of particles in one or more samples. Generally, the size and / or shape of the system may be configured to be portable and / or mountable on laboratory benches, tables, shelves, etc. In some variations, the system may comprise at least a first module, a second module, and a third module, which are housed within a housing and configured to characterize particle samples and / or set samples for characterization.

[0047] For example, such as Figure 1A and 1B As shown, the system (100) may include a housing (102) within which a first module (104), a second module (106), and a third module (108) may be located on a substrate (116). A plate drawer (e.g., a microporous plate drawer (110) of the second module (106)) may be configured to open and extend through the front wall (112) of the housing (102) to an open position, allowing a sample plate (114) to be inserted into the drawer (110), and retract into the housing (102) to a closed position, so that the sample plate (114) is placed within the housing (102). Once placed within the housing (102), multiple samples in the sample plate (114) may be analyzed by one or more of the modules of the system (100). Multiple indicator lights (111) may further be provided on the front wall (112) of the housing (102) to indicate the status of the system (e.g., various states). For example, a flashing white light indicates that the system is starting up; a solid white light indicates that the system has started up correctly and is ready for use; a flashing blue light indicates that the microplate drawer is being moved in / out or opened, the plate is still inside the system, and / or the system is still measuring; a solid red light indicates that the system is in an error state; a flashing red light indicates that the microplate drawer is obstructed; and a flashing yellow light indicates the self-test mode after startup.

[0048] A module of the system may be coupled to one or more different modules of the system via a mechanical connection (e.g., via a cable) or a wireless connection. The components of the system (e.g., housing, modules, sample plates) may be made of any suitable material (e.g., any suitable polymer material, metallic material, or glass material) or any suitable combination of materials.

[0049] Module 1 (Dynamic Light Scattering Module)

[0050] As previously described, a system for characterizing particles may include one or more processors, multiple samples, and a first module configured to measure light scattered by each of the multiple samples. One or more processors may be coupled to the first module (and / or other modules, such as a second or third module) via wired or wireless connections. Each of the multiple samples may contain one or more types of particles. Samples may contain any type of particles that scatter light when irradiated. Exemplary types of particles may include, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof.

[0051] The first module may include one or more light emitters, a single photodetector, and an actuator with a rotation axis. The one or more light emitters may contain a laser source. The laser source can be any laser source or configuration capable of providing monochromatic and / or polarized light. In some variations, the laser source may have a power of approximately 40 mW and produce a wavelength of approximately 660 nm. When multiple light emitters are used, they may contain the same or different laser sources. The one or more laser sources may be housed within the system's housing.

[0052] A single photodetector may include an optical fiber and be coupled to an actuator. The coupling between the photodetector and the actuator can be achieved using clamps, threads, keys, or other types of connections. In some variations, a single photodetector may be coupled to the actuator via a clamp, such that rotation of the actuator causes the photodetector to rotate about the actuator's axis of rotation. Rotation of the single photodetector about its axis of rotation allows it to receive light scattered by each sample at multiple scattering angles. The single photodetector can rotate on a sample plate holding multiple samples to detect scattered light at multiple scattering angles.

[0053] In some variations, a single photodetector can rotate about 1.0 degree to about 280 degrees (inclusive of all values ​​and subranges) around the rotation axis. In other words, the angular scanning of a single photodetector can be within the range of about 1.0 degree to about 280 degrees (inclusive of all values ​​and subranges). For example, a single photodetector can rotate about 1.0 degree, 5.0 degree, 10 degree, about 15 degree, about 20 degree, about 25 degree, about 30 degree, about 35 degree, about 40 degree, about 45 degree, about 50 degree, about 55 degree, about 60 degree, about 65 degree, about 70 degree, about 75 degree, about 80 degree, about 85 degree, about 90 degree, about 95 degree, about 100 degree, about 105 degree, about 110 degree, about 115 degree, about 120 degree, about 125 degree, and about 13 degrees around the rotation axis. 0 degrees, approximately 135 degrees, approximately 140 degrees, approximately 145 degrees, approximately 150 degrees, approximately 155 degrees, approximately 160 degrees, approximately 165 degrees, approximately 170 degrees, approximately 175 degrees, approximately 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 rotation axis, or rotate more than 280 degrees around the rotation axis. In other variations, rotation of the actuator between approximately 30 degrees and approximately 180 degrees also causes the single photodetector to rotate by the corresponding number of degrees.

[0054] The scattered light received by a single photodetector may include forward-scattered light and / or back-scattered light from multiple scattering angles. When forward scattering is received, the multiple scattering angles may be in the range of approximately 25 degrees to approximately 45 degrees (inclusive of all values ​​and subranges therein). For example, the forward scattering angles among the multiple scattering angles may be approximately 25 degrees, approximately 26 degrees, approximately 27 degrees, approximately 28 degrees, approximately 29 degrees, approximately 30 degrees, approximately 31 degrees, approximately 32 degrees, approximately 33 degrees, approximately 34 degrees, approximately 35 degrees, approximately 36 degrees, approximately 37 degrees, approximately 38 degrees, approximately 39 degrees, approximately 40 degrees, approximately 41 degrees, approximately 42 degrees, approximately 43 degrees, approximately 44 degrees, or approximately 45 degrees.

[0055] When backscattered light is received, multiple scattering angles can range from approximately 105 degrees to approximately 170 degrees (inclusive of all values ​​and subranges). For example, the backscattering angles among the multiple scattering angles can be approximately 105 degrees, approximately 106 degrees, approximately 107 degrees, approximately 108 degrees, approximately 109 degrees, approximately 110 degrees, approximately 111 degrees, approximately 112 degrees, approximately 113 degrees, approximately 114 degrees, approximately 115 degrees, approximately 116 degrees, approximately 117 degrees, approximately 118 degrees, approximately 119 degrees, approximately 120 degrees, approximately 121 degrees, approximately 122 degrees, approximately 123 degrees, approximately 124 degrees, approximately 125 degrees, approximately 126 degrees, approximately 127 degrees, approximately 128 degrees, approximately 129 degrees, approximately 130 degrees, approximately 131 degrees, 132 degrees, 133 degrees, approximately 134 degrees, approximately 135 degrees, approximately 170 ... 36 degrees, approximately 137 degrees, approximately 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.

[0056] The light emitter may be an optical fiber and may include a laser source (e.g., a monochromatic source), as described above. The system may include one or more light emitters. When multiple light emitters are used, they 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 in 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, such as... Figure 2A As shown, the first module (200) (configured to at least emit light and detect scattered light) may include a first light emitter (202) disposed above the sample plate (204) and a second light emitter (206) disposed below the sample plate (204). The illumination of the first light emitter and the second light emitter may be sequential (i.e., not simultaneous). Illumination of the first light emitter (202) of the sample in the sample plate (204) from the top may result in backscattering of light. Illumination of the second light emitter (206) of the sample in the sample plate (204) from the bottom may result in forward scattering of light.

[0057] The system may include a single photodetector configured to receive scattered light at multiple scattering angles, as previously stated. The single photodetector may be connected to a photon counter and 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 it rotates as the actuator rotates about its axis of rotation (e.g., its longitudinal axis). For example, see reference... Figure 2A and 2B A single photodetector (208) positioned above the sample plate (204) can be coupled to an actuator (210), which can be configured to rotate about a rotation axis (212). More specifically, the single photodetector (208) can be coupled to the actuator (210) via a clamp (214) on a swing arm (216) of the actuator (210). Figure 2C As shown, the actuator (210) can rotate the photodetector (208) in the direction of arrow (218). Rotation can be performed about 1.0 degree to about 280 degrees (inclusive of all values ​​and subranges) about the rotation axis (212). In some variations, the angle of the swing arm (216) can be moved in increments of about 1.8 degrees. This wide angular scanning allows for the detection and analysis of a larger amount of scattered light, and thus results in more accurate particle sizes within the sample. In one variation, rotating the photodetector and selecting illumination from a first light emitter (e.g., above the sample plate) and / or a second light emitter (e.g., below the sample plate) can be included in the process of setting the scattering angle for detecting light scattering from the sample. Irradiation from the light emitter can have an emitted beam axis parallel to the longitudinal axis from which it originates.

[0058] As further described below, the sample plate may include multiple cuvettes that hold small volumes of sample for characterization / analysis. A first module may be configured to adjust the position of the emitted beam axis such that the intersection (beam overlap) between the emitted beam axis and the detector beam axis is maintained. This adjustment may be performed using one or more actuators (e.g., stepper motors). The actuators may be adjusted manually or automatically based on instructions from one or more system processors. Given that any deformation of the sample plate (e.g., the sample plate surface) may distort the optical path of the emitted beam and / or the received beam (which contains light scattered at a specific scattering angle), it may be useful to restore the intersection of the emitted and received beams each time the photodetector moves (e.g., rotates) and / or the illumination direction changes (e.g., from above to below the sample plate, and vice versa), so that beam overlap occurs at the same location (this can help improve the accuracy of the obtained data).

[0059] For example, refer to Figure 3AWhen the photodetector rotates, it may be useful to restore the beam overlap between the emitted beam (300) and the received beam (302), so that the overlap forms a cone shape. Without restoring the beam overlap, the emitted beam (304) and the received beam (306) can form a hyperboloid, such as... Figure 3B As shown in the diagram. Defects in the pantograph and the facet of the photodetector can also cause deformation in the optical paths of the emitted and received beams. The first module may include a stage configured to move one or more light emitters in a horizontal plane in the X' and / or Y' directions, such that beam overlap can be restored. For example, refer to... Figure 3C The first module (308) may include a stage (310) including a first stage actuator (310a) and a second stage actuator (310b). The first stage actuator is configured to translate a light emitter (312) in the Y' direction (one emitter is disposed above the sample plate (314) and the other emitter is disposed below the sample plate (314). The second stage actuator is configured to translate the light emitter (312) in the X' direction.

[0060] The first module may further include a third stage actuator (e.g., a stepper motor) configured to translate the photodetector and / or light emitter in a vertical plane in the Z' direction, such that the intersection (beam overlap) of the emitted and received beams occurs at the same height in the plurality of cuvettes. For example, refer to Figure 4A The first module (400) may include a third stage actuator (408) configured to translate the first module (400) and associated single photodetector (402) and / or light emitter (404) (one emitter positioned above the sample plate (406) and one emitter positioned below the sample plate (406)) in a vertical direction (e.g., the Z' direction). Figure 4B and 4C As further detailed, the beam overlap (410) of the emitted beam (412) and the received beam (414) is maintained at the same height (H) in the cuvette (416). Figure 4B The image shows the beam overlap (410) when light is emitted from a light emitter (418) above the sample plate (420), and Figure 4C The beam overlap (410) is shown when light is irradiated from a light emitter (422) below the sample plate (420). In one variation, when the cuvette height is 0.7 mm, it may be useful to maintain the crossover point at a height of 0.35 mm in each of the multiple cuvettes.

[0061] Second module (sample plate positioning module)

[0062] Some variations of the system described herein may include a second module configured to move the sample plate to a location where light scattering and / or UV / Vis measurements are desired. For example, refer to... Figure 1B The second module (106) may include a microplate drawer (110) configured to receive a sample plate (114) therein and adjust the position of the sample plate (114) within the housing (102) to obtain light scattering and / or absorptivity data. The second module may be configured for manual or automatic translation. One or more second module actuators (e.g., motors) may be used to achieve sample plate positioning. For example, one or more second module actuators may be used to move the sample plate such that each cuvette is properly positioned for obtaining measurements via a single photodetector and / or for moving the plate such that each cuvette of the sample plate can be analyzed sequentially. When the position adjustment is automatic, the second module may be configured to receive instructions from one or more processors of the system based on information input by the user or based on, for example, data / information from system calibration, one or more types of particles sampled, cuvette size, etc. A single photodetector may rotate on the sample plate holding multiple samples to detect scattered light at multiple scattering angles.

[0063] The sample plates set within the second module can be disposable and configured in various ways, but are typically constructed to hold one or more samples. When the plate contains multiple samples, it can hold 2 to 96 samples. For example, the plate can hold 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 The sample plates can hold 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. In some variations, the sample plates can be designed for automated, high-throughput use (e.g., by employing a stackable design and incorporating barcodes for tracking). The sample plates can also be configured for manual pipetting of samples or be compatible with liquid handling robots that feed samples into the wells of the plate.

[0064] The sample plate may have a length ranging from about 11 cm to about 13 cm (inclusive of all values ​​and subranges therein). 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 range from about 7.0 cm to about 9.0 cm (inclusive of all values ​​and subranges therein). 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 materials, the sample plate may be made of polymers (e.g., thermoplastic polymers), plastics, metals, glass, or combinations thereof. In one variation, the sample plate may be made of acrylonitrile butadiene styrene (ABS).

[0065] In some variations, the sample plate may include multiple cuvettes in which a sample is contained to obtain light scattering and / or absorbance data. The multiple cuvettes may be mounted on a component configured as strips (also referred to herein as “chips” or “microfluidic chips”). The sample plate may contain one or more strips. For example, the sample plate may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 strips. In some variations, it may be advantageous for the sample plate to contain six strips. One or more strips may be pre-mounted on the substrate of the plate.

[0066] The strip may have both length and width. The length of the strip may range from about 7.0 cm to about 9.0 cm (inclusive of all values ​​and subranges 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 (inclusive of all values ​​and subranges 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, a strip with a length of about 8.1 cm and a width of about 1.8 cm may be useful. The strip may be made of any suitable material, including but not limited to polymers (e.g., thermoplastic polymers), plastics, glass, and combinations thereof. In one variation, the strip may be made of a cyclic olefin copolymer (COC). In another variation, the strip may be made of glass. In yet another variation, the strip may be formed from a material that makes it transparent.

[0067] Multiple wells configured to receive samples (e.g., input wells) may also be included on the strip. The strip may contain any number of wells. For example, the strip may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 wells. The wells may have a tapered 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 can be very small, ranging from about 0.1 μl to about 2.5 μl (inclusive of all values ​​and subranges therein). 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 can be, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. The size of one or more types of particles in a sample can range from about 0.3 nm to about 3,000 nm (inclusive of all values ​​and subranges therein).For example, the size of one or more particles can be 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 60 nm, approximately 65 nm, approximately 70 nm, approximately 75 nm, approximately 80 nm, approximately 85 nm, approximately 90 nm, etc. nm, approximately 95 nm, approximately 100 nm, approximately 150 nm, approximately 200 nm, approximately 250 nm, approximately 300 nm, approximately 350 nm, approximately 400 nm, approximately 450 nm, approximately 500 nm, approximately 550 nm, approximately 600 nm, approximately 650 nm, approximately 700 nm, approximately 750 nm, approximately 800 nm, approximately 850 nm, approximately 900 nm, approximately 1,000 nm, approximately 1,500 nm, approximately 2,000 nm, approximately 2,500 nm, or approximately 3,000 nm.

[0068] The strip can be configured such that a sample placed in the well flows through at least one channel (e.g., at least one microchannel) to one or more cuvettes for analysis. The cuvettes can have a path length in the range of about 0.1 mm to about 0.7 mm (inclusive of all values ​​and subranges therein). For example, the path length can 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.

[0069] In some variations, the sample plate can be as follows: Figure 5A Configure as shown in the image. (Refer to...) Figure 5A The sample plate (500) may contain multiple strips on a substrate (504), such as six strips (502). Figure 5B A top view of one of the six strips (502) is provided separately, showing the configuration of the multiple holes (504) disposed on the strip (502). Figure 5C A magnified view of the framed area (E) is provided. Figure 5CIn the diagram, one of the multiple holes (504) is shown as fluidly connected to at least one cuvette (506) via a channel (508). Although the cuvette (506) is shown as having a circular-rhomboid shape, other shapes are possible. For example, the cuvette can be shaped as a circle, oval, triangle, square, or rectangle. Reference Figures 6A to 6D When placed into hole (600) Figure 6A After that, the sample (yellow) can be drawn into the channel (602) by capillary force. Figure 6B To avoid evaporation, the empty cuvettes (604, 606) are then read. Figure 6C A small vacuum pressure (e.g., about 20 mm Hg) from one or more pumps in the system can then be applied to the vent (608) to move the sample into cuvettes (604) and / or (606) and to perform light scattering measurements. Figure 6D The pump can be housed inside the system housing or located outside the system housing.

[0070] Module 3 (Spectrometer Module)

[0071] In some variations, the system described herein may include a third module configured to perform UV / Vis spectroscopy. For example, see reference... Figure 1A and 1B The third module (108) can be positioned adjacent to the second module (106) and the first module (104) within the housing (102). Thus, in these variations, the system can measure light scattering at multiple angles and the absorbance of the sample as a function of wavelength, in the range of, for example, 230 nm to 750 nm. Based on the measured UV / Vis absorption spectra, the number and mass of particles can be calculated. Dynamic light scattering measurements obtained at multiple angles using a single rotating photodetector of the first module can be used for particle size determination and / or aggregation detection as described herein.

[0072] In use, absorbance measurements can be obtained from the sample after it has been transferred to one or more cuvettes. Therefore, return to the reference. Figures 6A to 6D Once the sample is moved into cuvettes (604) and / or (606), absorbance measurements can be obtained using the spectrometer in the third module. Absorbance measurements can be obtained before or after light scattering measurements (described above). In some instances, the system can simultaneously measure UV / Vis absorption through the cuvette during the pressure-driven transport of the sample from the channel to the cuvette. This allows the system to monitor the cuvette filling behavior in addition to analyzing the spectral absorbance of the sample.

[0073] The system may contain any number of modules or combinations of modules, but will typically 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 one or more particle sizes and / or particle size distributions; a second module for appropriately setting up sample plates and cuvettes for performing measurements, calibrations, and / or obtaining other data; and a third module for measuring the absorbance of one or more samples. The first, second, and third modules may be housed within the system's housing, such as... Figure 1A and 1B As shown in the illustration. Other system components (e.g., vacuum pumps, one or more laser sources, and processors / processing units) may also be included within the housing. However, in some variations, one or more processors / processing units may be located outside the housing (e.g., in a computer) and coupled to the system module, for example, via wired or wireless communication. As previously mentioned, data from light scattered at multiple (e.g., different) angles can be combined and analyzed by one or more processors to determine a single result that may be angle-independent. In some variations, one or more processors may employ algorithms configured to determine particle size using data and / or signals correlated with one or more of the following: sample temperature, sample viscosity, hydrodynamic diameter of the particles, scattered light intensity, the ratio of scattered light intensity to incident light intensity at a specific angle, the scattered light angle, and the rate of change of scattered light intensity (e.g., fluctuations in scattered light intensity over time). In some variations, one or more processors may include correlators configured to analyze particle size based on data and / or intensity of the various signals described herein.

[0074] Additionally, the system may include a display configured to show / present various types of information / data to the user, such as system status, alarms, 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 mounted on a portion of the housing or may include a monitor coupled to the system. The display may also include or act as a user interface through which the user can control one or more modules and / or components of the system. In this example, the user interface may include a touchscreen. 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.

[0075] One or more processors can be configured to control the operation of system modules, other system components, and their interactions. Thus, one or more processors can be configured to execute instructions for setting and quantifying the size of one or more particles or other substances within a sample. For example, one or more processors of the system can be configured to determine the size of one or more types of particles in a sample based on data from scattered light. As previously mentioned, the size of one or more types of particles in the sample can range from about 0.3 nm to about 3,000 nm (inclusive of all values ​​and subranges therein). One or more processors can be configured to receive multiple signals from a single photodetector, each of which can represent scattered light at a specific angle. The data from light scattered at multiple (e.g., different) angles can then be combined and analyzed by one or more processors to determine a single result that is angle-independent, as further described below.

[0076] method

[0077] This document also describes methods for characterizing particles. Exemplary particles include, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, and / or small molecules. Generally, the method may include: irradiating multiple samples with light from one or more light emitters, each of the multiple samples containing one or more types of particles; rotating a photodetector about a rotation axis within multiple scattering angles; detecting the light scattered by the particles within the multiple scattering angles; and obtaining light scattering measurements for each of the multiple scattering angles. The rotation of the photodetector may occur above the multiple samples (i.e., above a sample plate). As mentioned above, data from the light scattered at the multiple scattering angles can then be combined and analyzed by one or more processors to determine individual results that are independent of angle.

[0078] One or more light emitters may include a laser source. It should be 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 these variations, the first light emitter may be positioned above multiple samples, and the second light emitter may be positioned below multiple samples. When two light emitters are used (e.g., a first light emitter above the sample plate and a second light emitter below the sample plate), the illumination of each light emitter is typically sequential. In other words, the first light emitter may illuminate each of the multiple samples, followed by the second light emitter, or vice versa.

[0079] Rotation can be achieved using an actuator that, when rotated about its axis of rotation (e.g., its longitudinal axis), causes the photodetector to rotate. As the photodetector rotates to obtain scattered light at multiple angles, the rotation angle can range from about 1 degree to about 280 degrees (inclusive of all values ​​and sub-ranges) about the actuator's axis of rotation. For example, a single photodetector can rotate about 1.0 degree, 5.0 degree, 10 degree, about 15 degrees, about 20 degrees, about 25 degrees, about 30 degrees, about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees, about 90 degrees, about 95 degrees, about 100 degrees, about 105 degrees, about 110 degrees, about 115 degrees, about 120 degrees, about 125 degrees, and about 13 degrees about the rotation axis. 0 degrees, approximately 135 degrees, approximately 140 degrees, approximately 145 degrees, approximately 150 degrees, approximately 155 degrees, approximately 160 degrees, approximately 165 degrees, approximately 170 degrees, approximately 175 degrees, approximately 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 or more around the rotation axis. In other variations, rotation of the actuator between approximately 30 degrees and approximately 180 degrees also causes a single photodetector to rotate by the corresponding number of degrees. This wide-angle scanning allows for the detection and analysis of a larger amount of scattered light, resulting in more accurate particle sizes within the sample.

[0080] Both forward-scattered and back-scattered light can be detected by a single rotating photodetector. Forward scattering can be detected at multiple scattering angles within a range of approximately 25 degrees to approximately 45 degrees (inclusive of all values ​​and subranges). For example, the forward scattering angles among the multiple scattering angles can be approximately 25 degrees, approximately 26 degrees, approximately 27 degrees, approximately 28 degrees, approximately 29 degrees, approximately 30 degrees, approximately 31 degrees, approximately 32 degrees, approximately 33 degrees, approximately 34 degrees, approximately 35 degrees, approximately 36 degrees, approximately 37 degrees, approximately 38 degrees, approximately 39 degrees, approximately 40 degrees, approximately 41 degrees, approximately 42 degrees, approximately 43 degrees, approximately 44 degrees, or approximately 45 degrees.

[0081] Backscattering can be detected at multiple scattering angles within the range of approximately 105 degrees to approximately 170 degrees (inclusive of all values ​​and subranges). For example, the backscattering angles among the multiple scattering angles can be approximately 105 degrees, approximately 106 degrees, approximately 107 degrees, approximately 108 degrees, approximately 109 degrees, approximately 110 degrees, approximately 111 degrees, approximately 112 degrees, approximately 113 degrees, approximately 114 degrees, approximately 115 degrees, approximately 116 degrees, approximately 117 degrees, approximately 118 degrees, approximately 119 degrees, approximately 120 degrees, approximately 121 degrees, approximately 122 degrees, approximately 123 degrees, approximately 124 degrees, approximately 125 degrees, approximately 126 degrees, approximately 127 degrees, approximately 128 degrees, approximately 129 degrees, approximately 130 degrees, approximately 131 degrees, 132 degrees, 133 degrees, approximately 134 degrees, approximately 135 degrees, and approximately 170 degrees. 36 degrees, approximately 137 degrees, approximately 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.

[0082] In some variations, the method may further include adjusting the position of the emission beam axis such that when light scattering is measured at multiple scattering angles and / or when the illumination direction changes (e.g., from the top to the bottom of the plate, and vice versa), the intersection point (beam overlap) between the emission beam axis and the detector beam axis in each cuvette is restored. 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, which can be restored such that it is the same as or substantially the same as the first beam overlap, such as... Figure 3A As shown in the diagram. It is also possible to recover light scattering measured at subsequent scattering angles (e.g., the third, fourth, fifth, sixth, seventh, and eighth scattering angles) such that the beam overlap is the same or substantially the same, causing the beam overlap to form a conical rather than a hyperboloid shape, as... Figure 3A (conical) and Figure 3B The comparison between (hyperboloids) is shown. The position of the emitted beam axis can be adjusted manually or automatically according to instructions from one or more processors. It may be necessary to restore beam overlap to account for factors such as defects and / or deformations of the strip surface in the arm of the rotary actuator or the facet of the photodetector, which may distort the optical paths of the emitted beam and the received beam containing scattered light.

[0083] Adjustments to restore the emitted beam axis to restore beam overlap (e.g., automatic adjustment) can be based on an algorithm (e.g., processing / method steps) run by one or more processors, which includes identifying a maximum intensity measurement. The algorithm may include one or more preliminary positioning steps. For example, and as... Figures 7A to 7D As shown, one or more preliminary positioning steps may include moving one or more light emitters to theoretical positions on the hyperboloid, such as the minimum circle, during system calibration. Figure 7A Next, the algorithm may include one or more fine-tuning steps. In some variations, fine-tuning may include steps such as moving the light emitter (2D scanning) and creating a 2D map of the intensity to find the desired overlap (e.g., maximum intensity), as shown below. Figure 7B As shown in the image. Reference Figure 7A and 7B As the light moves along line A, the intensity can remain constant; line A is the bisector of the line between the light emitter and the photodetector. Alternatively, fine positioning may further include the step of scanning the intensity along line A to find the maximum intensity. For example, Figure 7C The intensity measured along line A is shown. Given that the intensity along line A is known to be Gaussian, measuring a finite number of points (e.g., four, five, six, or seven points) is sufficient. For each point (indicated by five black dots), as... Figure 7D As shown, the points can be fitted and / or their positions refined on a Gaussian curve, the maximum value of the curve can be identified, and the intensity at the maximum value can be measured.

[0084] The method may also include maintaining beam overlap at the same height in each cuvette while obtaining light scattering measurements from multiple cuvettes. In some variations, the same height may be maintained using an actuator, such as the third stage actuator described above, which may be configured to translate the photodetector and / or light emitter in a vertical plane in the Z' direction such that the intersection point (beam overlap) of the emitted and received beams occurs at the same height in the multiple cuvettes. The steps involved in maintaining beam overlap may be as follows: Figures 4A to 4C As shown in the image. Reference. Figure 4A The first module (400) may include a third stage actuator (408) configured to translate the first module (400) and associated single photodetector (402) and / or light emitter (404) (one emitter positioned above and one below the sample plate (406)) in a vertical direction (e.g., the Z' direction). Figure 4B and 4C As further detailed, the beam overlap (410) of the emitted beam (412) and the received beam (414) is maintained at the same height (H) in the cuvette (416). Figure 4BThe image shows the beam overlap (410) when light is emitted from a light emitter (418) above the sample plate (420), and Figure 4C The beam overlap (410) is demonstrated when light is irradiated from a light emitter (422) below the sample plate (420). In one variation, when the cuvette height is 0.7 mm, it may be useful to maintain the crossover point (beam overlap) at a height of 0.35 mm in each of the multiple cuvettes. The beam overlap can be adjusted manually or automatically according to instructions from one or more processors.

[0085] Alternatively, the method may include obtaining absorbance data (e.g., UV / Vis data) from multiple samples. For example, multiple samples can be irradiated with light having wavelengths in the range of about 190 nm to about 900 nm, and the absorbance can be measured. The absorbance data can be obtained using a module of the system (e.g., the second module described above). The absorbance data can be obtained before or after obtaining the light scattering data.

[0086] Particle characterization can be performed on small sample volumes ranging from about 0.5 μl to about 2.5 μl (inclusive of all values ​​and subranges therein). For example, sample volumes can 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 variant, multiple samples have a volume of about 2 μl. In another variant, multiple samples are approximately 2 μl or less in volume.

[0087] Determining the size of one or more types of particles in a sample can be accomplished by one or more processors of a system configured to analyze data from scattered light. The size of one or more types of particles in a sample can range from about 0.3 nm to about 3,000 nm (inclusive of all values ​​and subranges therein). For example, the size of one or more particles can be 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 60 nm, approximately 65 nm, approximately 70 nm, approximately 75 nm, approximately 80 nm, approximately 85 nm, approximately 90 nm, etc. The particle sizes are approximately 95 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm, 900 nm, 1,000 nm, 1,500 nm, 2,000 nm, 2,500 nm, or 3,000 nm. The particles may include, but are not limited to, nanoparticles, DNA, RNA, viruses, proteins, polymers, small molecules, and combinations thereof, as previously mentioned herein.

[0088] In some variations, one or more processors may combine and analyze data from light scattered at multiple (e.g., different) angles to determine a single result that is angle-independent. For example, one or more processors may employ algorithms configured to determine particle size using data and / or signals correlated with one or more of the following: sample temperature, sample viscosity, hydrodynamic diameter of the particles, scattered light intensity, the ratio of scattered light intensity to incident light intensity at a specific angle, the scattered light angle, and the rate of change of scattered light intensity (e.g., fluctuations in 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 the various signals described above.

[0089] In some variations, once signals are obtained from scattered light detected by a single photodetector at multiple scattering angles, one or more processors can combine and analyze the data from the light scattered at multiple (e.g., different) angles to determine (e.g., for calculation) a single result (e.g., a single solution) characterizing the particles (e.g., particle size and / or particle size distribution) in the sample. The single solution can be angle-independent. In one variation, determining the single solution involves an absolute correlation function that satisfies all expectations at once; the single solution is represented by the following equation:

[0090]

[0091] Where R(q) is the ratio of the scattering intensity to the Rayleigh scattering intensity at each angle (the Rayleigh ratio at angle q), and the correlation function is the square root of the correlation function multiplied by R(q). The correlation function can be transformed, including adjustments for different scattering angles, temperatures, and viscosities (e.g., by rescaling the x-axis).

[0092] In some variations, one or more processors can use absolute correlation data obtained from light scattered at multiple angles to create a graphical representation of the absolute correlation function, such as a 2D representation or model. For a single particle, the 2D graphical representation could be a model with values ​​at x and q. 2 A plane with a slope tilted in the direction of scattering. Therefore, for a mixture of particles, a 2D graphical representation can contain the sum of such planes (e.g., planes are represented collectively). In one variation, the 2D representation can help determine the size of one or more types of particles in each sample based on the shape of the shape, which is a function of the absolute correlation of multiple scattering angles, such as... Figures 8A to 8C As shown in the figure. For example, when the sample contains relatively small particles (e.g., 10 nm), the shape of the absolute correlation function can be as follows. Figure 8A As shown in the figure, all planes are parallel, and there is rapid decay in the x-direction and in the q-direction. 2 There is no attenuation in the direction. When the sample contains relatively large particles (e.g., 100 nm), the shape of the absolute correlation function can be as follows: Figure 8B As shown in the figure, all the planes are parallel, but there is a slow decay in the x-direction and at q 2 There is rapid decay in the direction. When the sample contains a mixture of particles (e.g., 10 nm and 100 nm), the shape of the absolute correlation function can be as follows: Figure 8C As shown in the figure, the planes are not parallel and fan out in the x-direction.

[0093] Although the foregoing variations have been described in detail with reference to illustrations and examples for clarity and understanding, it will be apparent that certain changes and modifications may be practiced and are intended to fall within the scope of the appended claims. Furthermore, it should be understood that the components and features of the systems and apparatuses described herein can be used in any combination. The description of certain elements or features with respect to particular figures is not intended to be restrictive, nor should it be construed as implying that such elements cannot be used in combination with any other described elements. For all variations described herein, the steps of the method may be performed in any order. Some steps are optional, such that each step of the method may be omitted.

Claims

1. A system for characterizing particles, comprising: One or more processors; Multiple samples, wherein each of the multiple samples comprises one or more types of particles; and A first module, configured to measure the light scattered by each sample, includes: One or more light emitters; An actuator having a rotation axis; A single photodetector is configured to rotate about the axis of rotation and receive light scattered by each sample at multiple scattering angles.

2. The system of claim 1, further comprising a sample plate configured to hold the plurality of samples.

3. The system according to claim 2, wherein the sample plate comprises a plurality of cuvettes.

4. The system of claim 3, wherein the first module includes one or more stage actuators configured to adjust the position of the emission beam axis such that when the photodetector receives light scattered at each of the plurality of scattering angles, the intersection point (beam overlap) between the emission beam axis and the detector beam axis is recovered in each sample.

5. The system of claim 4, wherein the one or more stage actuators are configured to automatically adjust the position of the emission beam axis.

6. The system of claim 4, wherein the one or more stage actuators are further configured to adjust the position of the first module such 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.

7. The system of claim 2, further comprising a second module configured to automatically adjust the position of the sample plate according to instructions from the one or more processors.

8. The system of claim 1, wherein the single optical detector comprises an optical fiber.

9. The system of claim 1, wherein the single photodetector is configured to rotate about 1.0 degree to about 280 degrees about the rotation axis.

10. The system of claim 1, wherein the single photodetector is configured to receive forward light scattering from the plurality of scattering angles.

11. The system of claim 10, wherein the plurality of scattering angles are in the range of about 25 degrees to about 45 degrees.

12. The system of claim 1, wherein the single photodetector is configured to receive backscattered light from the plurality of scattering angles.

13. The system of claim 12, wherein the plurality of scattering angles are in the range of about 105 degrees to about 170 degrees.

14. The system of claim 1, further comprising a third module configured to irradiate the plurality of samples with light having a wavelength in the range of about 190 nm to about 900 nm, and to measure the absorptivity of the plurality of samples.

15. The system of claim 1, wherein the one or more light emitters comprise a laser light source.

16. The system of claim 1, wherein the one or more optical emitters comprise a first optical emitter and a second optical emitter.

17. The system of claim 16, wherein the first light emitter is disposed above the sample plate and the second light emitter is disposed below the sample plate.

18. The system of claim 1, wherein the one or more types of particles include nanoparticles, DNA, RNA, viruses, proteins, polymers, or small molecules.

19. The system of claim 1, wherein each of the plurality of samples is about 2 μl or less.

20. The system of claim 3, wherein the sample plate comprises a plurality of components, each of the plurality of components being configured as a strip having a length and a width.

21. The system of claim 20, wherein the plurality of components comprises six strips.

22. The system of claim 20, wherein the strip includes a plurality of wells configured to receive the plurality of samples.

23. The system of claim 22, wherein each of the plurality of holes is coupled via a channel to a corresponding cuvette from the plurality of cuvettes.

24. The system of claim 3, wherein each of the plurality of cuvettes has a path length in the range of about 0.1 mm to about 0.7 mm.

25. The system of claim 24, wherein the path length is approximately 0.1 mm.

26. The system of claim 24, wherein the path length is approximately 0.7 mm.

27. The system of claim 1, wherein the one or more processors are configured to determine the size of the one or more types of particles in the sample.

28. The system of claim 27, wherein the size of the one or more types of particles in the sample is in the range of about 0.3 nm to about 3,000 nm.

29. A system comprising: The first module includes a photodetector configured to rotate above a sample plate containing multiple samples and detect light scattered from each sample at multiple scattering angles due to its rotation. and The second module is configured to measure the absorbance of each of the plurality of samples. Each of the plurality of samples comprises one or more types of particles.

30. The system of claim 29, further comprising one or more processors configured to determine particle size or particle size distribution from the scattered light detected at the plurality of scattering angles.

31. A method for characterizing particles, comprising: Irradiate multiple samples with light from one or more light emitters, wherein each of the multiple samples contains one or more types of particles; The photodetector is rotated around the rotation axis within multiple scattering angles; Detect the light scattered by the particles within the plurality of scattering angles; and Obtain light scattering measurements for each of the plurality of scattering angles.

32. The method of claim 31, further comprising adjusting the position of the emission beam axis such that when the photodetector receives light scattered at each of the plurality of scattering angles, the intersection point (beam overlap) between the emission beam axis and the detector beam axis is recovered in each sample.

33. The method of claim 32, further comprising adjusting the position of the intersection point (beam overlap) between the emitted beam axis and the detector beam axis such that the beam overlap is maintained at the same height in the plurality of cuvettes.

34. The method of claim 32, further comprising automatically adjusting the position of the axis of the emitted beam.

35. The method of claim 31, further comprising automatically adjusting the position of the sample plate according to instructions from one or more processors.

36. The method of claim 33, wherein the instructions include one or more preliminary positioning steps.

37. The method of claim 36, wherein the one or more preliminary positioning steps include moving the one or more light emitters to the smallest circle on the hyperboloid.

38. The method of claim 36, further comprising one or more fine positioning steps.

39. The method of claim 38, wherein the one or more fine positioning steps include determining the maximum intensity of the light scattered by each sample at the plurality of scattering angles.

40. The method of claim 31, wherein the optical detector comprises an optical fiber.

41. The method of claim 31, wherein the photodetector is configured to rotate about 1 degree to about 280 degrees about the rotation axis.

42. The method of claim 31, wherein the photodetector detects forward light scattering from the plurality of scattering angles.

43. The method of claim 42, wherein the plurality of scattering angles are in the range of about 25 degrees to about 45 degrees.

44. The method of claim 31, wherein the photodetector detects backscattered light from the plurality of scattering angles.

45. The method of claim 44, wherein the plurality of scattering angles are in the range of about 105 degrees to about 170 degrees.

46. ​​The method of claim 31, wherein the irradiation is performed using a laser light source.

47. The method of claim 31, wherein the one or more optical emitters comprise a first optical emitter and a second optical emitter.

48. The method of claim 47, wherein the first light emitter is disposed above the plurality of samples, and the second light emitter is disposed below the plurality of samples.

49. The method of claim 31, wherein the one or more types of particles include nanoparticles, DNA, RNA, viruses, proteins, polymers, or small molecules.

50. The method of claim 31, wherein each of the plurality of samples is about 2 μl or less.

51. The method of claim 31, further comprising using one or more processors to determine the size of the one or more types of particles in the sample based on the scattered light at the plurality of scattering angles.

52. The method of claim 51, wherein the size of the one or more types of particles in the sample is in the range of about 0.3 nm to about 3,000 nm.

53. The method of claim 51, wherein one or more processors determine the size of the one or more types of particles in each sample based on the shape of the absolute correlation function of the plurality of angles.

54. The method of 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 obtained light scattering measurements for the plurality of scattering angles.

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