Antibody concentration measurement, titer module, and liquid storage module for cell analysis system

Abstract

A system for measuring an antibody concentration in a sample includes a polarizer holder holding a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within a container to pass in a first direction. The second detection polarizer restricts fluorescence emitted from within the container to pass in a second direction. The second direction is perpendicular to the first direction. The system includes a detector for measuring fluorescence emitted in a first direction and fluorescence emitted in a second direction. The system rotates the polarizer holder in an alternating direction about an axis of rotation such that the first detection polarizer and the second detection polarizer are alternately placed in an optical path of fluorescence emitted from within the container.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Publication Number (43) Publication Date (21) Application Number 202480043356.5 (22) Application Date 2024.07.19 (30) Priority Data 63 / 514,929 2023.07.21 US (85) PCT International Application Entering National Phase Date 2025.12.26 (86) PCT International Application Application Data PCT / US2024 / 038864 2024.07.19 (87) PCT International Application Publication Data WO2025 / 024321 EN 2025.01.30 (71) Applicant: Beckman Coulter Ltd. Address: USA (72) Inventors: Jordan Bielefeld, Matthew Freeman, Allen Gustafson, Todd Halvorsen, Julie Rutti, Timothy Reed (74) Patent Agency: Beijing J&J Intellectual Property Agency Co., Ltd. 11227 Patent Attorney: Gao Yan (51) Int.Cl. G01N 21 / 64 (2006.01) G01N 33 / 53 (2006.01) (54) Invention Title: Antibody Concentration Measurement, Titer Module and Liquid Storage Module for Cell Analysis System (57) Abstract: A system for measuring antibody concentration in a sample, the system comprising: a polarizer holder holding a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within a container to pass through in a first direction. The second detection polarizer restricting fluorescence emitted from within a container to pass through in a second direction. The second direction is perpendicular to the first direction. The system includes a detector for measuring fluorescence emitted along the first direction and fluorescence emitted along the second direction. The system rotates a polarizer holder about a rotation axis in alternating directions, such that a first detection polarizer and a second detection polarizer are alternately positioned in the optical path of fluorescence emitted from within the container. Claims 3 pages, Description 23 pages, Drawings 38 pages, CN 121399448 A 2026.01.23 CN 1 21 39 94 48 A 1. A system for measuring antibody concentration in a sample, the system comprising: a light source; an excitation polarizer that polarizes light emitted from the light source along a first direction; a container configured to receive light polarized along the first direction; a polarizer holder holding a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within the container to pass through in the first direction, and the second detection polarizer restricting fluorescence emitted from within the container to pass through in a second direction perpendicular to the first direction;The system comprises: a detector for measuring fluorescence emitted along a first direction and fluorescence emitted along a second direction; and a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: rotate the polarizer holder about a rotation axis in alternating directions such that the first detection polarizer and the second detection polarizer are alternately positioned in the optical path of the fluorescence emitted from the container. 2. The system of claim 1, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to: rotate the polarizer holder to position the first detection polarizer in the optical path of the fluorescence emitted from the container; and rotate the polarizer holder to position the second detection polarizer in the optical path of the fluorescence emitted from the container. 3. The system of claim 1, further comprising: an optical plate having a groove terminating in a first stop and a second stop; and wherein the polarizer holder includes a pin that, when the polarizer holder rotates about the rotation axis, displaces within the groove of the optical plate. 4. The system of claim 3, wherein the optical plate further comprises a first magnet positioned next to the first stop in the groove and a second magnet positioned next to the second stop in the groove; and wherein the pin comprises a ferromagnetic material, and the pin is attracted to the first magnet when positioned next to the first stop at the end of a first rotational stroke, and is attracted to the second magnet when positioned next to the second stop at the end of a second rotational stroke. 5. The system of claim 3, wherein the groove is curved along the rotation axis. 6. The system of claim 1, wherein the polarizer holder comprises a first side surface and a second side surface perpendicular to the first side surface, the first side surface including a first recess for receiving the first detection polarizer, and the second side surface including a second recess for receiving the second detection polarizer. 7. The system of claim 6, wherein the polarizer holder further comprises a first biasing element attached to the first side surface to secure the first detection polarizer in the first recess, and a second biasing element attached to the second side surface to secure the second detection polarizer in the second recess. 8. The system according to claim 7, wherein when the first detection polarizer and the second detection polarizer are respectively placed in the first recess and the second recess, the first detection polarizer and the second detection polarizer protrude beyond the first side surface and the second side surface, and the first biasing element and the second biasing element... (Claim 1 / 3 page 2 CN 121399448)A. A bend is made around the first and second detection polarizers to secure the first and second detection polarizers in place. 9. The system of claim 8, wherein the first and second bias elements include cutouts to increase the curvature around the first and second detection polarizers. 10. The system of claim 6, wherein the polarizer holder further includes a curved surface connecting the first and second side surfaces, the curved surface having holes aligned with holes in the first and second recesses, thereby allowing the fluorescence emitted from the container to pass through the polarizer holder and reach the detector. 11. The system of claim 10, further comprising: an optical cover that at least partially accommodates the detector and the polarizer holder, the optical cover including a curved wall aligned with the curved surface of the polarizer holder to reduce the space between the polarizer holder and the curved wall. 12. The system of claim 1, further comprising: a working platform; a liquid storage module mounted on the working platform, the liquid storage module comprising: a base having an internal volume for receiving an internal reservoir; a main cover connected to the base, the main cover including a port providing access to the internal reservoir; a secondary cover attached to the main cover, the secondary cover being configured to seal the port on the main cover; a second motor attached to the secondary cover; and a dispensing system configured to move probes above the working platform along three mutually perpendicular axes; and wherein, when the instruction is executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to: operate the second motor to open the secondary cover, thereby allowing the probes of the dispensing system to be inserted through the port on the main cover to reach the bottom of the internal reservoir to draw liquid contained in the internal reservoir; and when the probes are removed from the port, operate the second motor to close the secondary cover to seal the port on the main cover. 13. A polarizer holder for a cell analysis system, the polarizer holder comprising: a first side surface; a second side surface perpendicular to the first side surface; a curved surface connecting the first side surface and the second side surface; a first detection polarizer coupled to the first side surface, the first detection polarizer restricting emitted fluorescence to pass in a first direction; and a second detection polarizer coupled to the second side surface, the second detection polarizer restricting emitted fluorescence to pass in a second direction; the polarizer holder being configured to rotate about a rotation axis such that the first detection polarizer and the second detection polarizer are alternately positioned in the optical path of the light. 14. The polarizer holder of claim 13, further comprising:A first recess in the first side surface, wherein the first detection polarizer is mounted in the first recess; and a second recess in the second side surface, wherein the second detection polarizer is mounted in the second recess. Claims 2 / 3, page 3, CN 121399448 A 15. The polarizer holder of claim 14, further comprising: a first biasing element attached to the first side surface to secure the first detection polarizer in the first recess; and a second biasing element attached to the second side surface to secure the second detection polarizer in the second recess. Claims 3 / 3 Page 4 CN 121399448 A Cross-Reference to Related Applications for Antibody Concentration Measurement, Titer Module, and Liquid Storage Module for Cell Analysis Systems

[0001] This application was filed as a PCT International Application on July 19, 2024, and claims the benefit and priority of U.S. Application No. 63 / 514,929, filed on July 21, 2024, entitled “ANTIBODY CONCENTRATION MEASUREMENT, TITER MODULE, AND LIQUID STORAGE MODULE FOR CELL ANALYSIS SYSTEM,” the disclosure of which is incorporated herein by reference in its entirety. Background Art

[0002] An antibody is a large Y-shaped protein that the immune system uses to recognize and neutralize foreign substances, such as pathogenic bacteria and viruses. Antibodies recognize unique molecules of pathogens (called antigens). Each tip of a Y-shaped antibody contains a complementary site that matches an epitope on the antigen, enabling the antibody to bind to the antigen. Using this binding mechanism, antibodies can tag microorganisms or infected cells for attack by other parts of the immune system, or can directly neutralize microorganisms or infected cells. Immunoglobulin G (IgG) is the most common type of antibody found in circulating blood. The concentration of antibodies in cell samples can be measured by fluorescence polarization. Summary of the Invention

[0003] Generally, this disclosure relates to measuring the concentration of antibodies in cell samples by fluorescence polarization. In one possible configuration, a measurement cycle is performed to reduce errors that may be caused by changes in signal intensity and noise during fluorescence polarization. Various aspects are described in this disclosure, including but not limited to the following aspects.

[0004] One aspect relates to a method of fluorescence polarization for measuring the concentration of antibodies in a sample, the method comprising: performing a measurement cycle, measuring a first set of sampling phases of fluorescence emitted along a first direction and a second set of sampling phases of fluorescence emitted along a second direction, at least one sampling phase of fluorescence emitted along the second direction occurring during fluorescence emitted along the first direction.Between sampling phases; calculating a first value for a first set of sampling phases; calculating a second value for a second set of sampling phases; and determining the antibody concentration based on a function of the first and second values.

[0005] On the other hand, a fluorescence polarization system for measuring antibody concentration in a sample is provided, the system comprising: a light source; a first polarization filter that polarizes light emitted from the light source along a first direction; a container containing a sample mixed with a fluorescent polarizing reagent, the container receiving light polarized along the first direction; movable second polarization filters and third polarization filters, the second polarization filter restricting fluorescence emitted from within the container to pass through in the first direction, and the third polarization filter restricting fluorescence emitted from within the container to pass through in a second direction substantially perpendicular to the first direction; a detector for... The system measures fluorescence emitted along a first direction and fluorescence emitted along a second direction; and a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: perform a measurement cycle; measure a first set of sampling phases of fluorescence emitted along the first direction and a second set of sampling phases of fluorescence emitted along the second direction, wherein at least one sampling phase of fluorescence emitted along the second direction occurs between sampling phases of fluorescence emitted along the first direction; calculate a first value of the first set of sampling phases; calculate a second value of the second set of sampling phases; and determine an antibody concentration based on a function of the first and second values.

[0006] On the other hand, a method for measuring the concentration of an antibody in a cell sample by fluorescence polarization is disclosed, the method comprising: performing a measurement cycle of fluorescence emitted from a cell sample mixed with a fluorescence polarizing reagent, the measurement cycle comprising the following measurement sequence: (1) measuring a first sampling phase of fluorescence emitted in a first direction before the midpoint of the measurement cycle; (2) measuring a second sampling phase of fluorescence emitted in a second direction before the midpoint of the measurement cycle, the second direction being perpendicular to the first direction; (3) measuring a third sampling phase of fluorescence emitted in the second direction after the midpoint of the measurement cycle; and (4) measuring a fourth sampling phase of fluorescence emitted in the first direction after the midpoint of the measurement cycle; calculating a first average value of the first sampling phase and the fourth sampling phase; calculating a second average value of the second sampling phase and the third sampling phase; and determining the antibody concentration based on a function of the first average value and the second average value.

[0007] On the other hand, a fluorescence polarization system for measuring the concentration of antibody proteins in a cell sample is provided. This system includes a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: perform a measurement cycle of fluorescence emitted from the cell sample, the measurement cycle including the following measurements...The sequence is as follows: (1) measuring a first sampling phase of fluorescence emitted along a first direction before the midpoint of the measurement period; (2) measuring a second sampling phase of fluorescence emitted along a second direction before the midpoint of the measurement period, the second direction being perpendicular to the first direction; (3) measuring a third sampling phase of fluorescence emitted along the second direction after the midpoint of the measurement period; and (4) measuring a fourth sampling phase of fluorescence emitted along the first direction after the midpoint of the measurement period; calculating a first average of the first sampling phase and the fourth sampling phase; calculating a second average of the second sampling phase and the third sampling phase; and determining the antibody concentration based on a function of the first average and the second average.

[0008] On the other hand, a fluorescence polarization system for measuring the concentration of immunoglobulin G (IgG) in a sample is provided. This system includes: a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: perform a measurement cycle; measure a first set of sampling phases of fluorescence emitted along a first direction and a second set of sampling phases of fluorescence emitted along a second direction substantially perpendicular to the first direction; at least one sampling phase of fluorescence emitted along the second direction occurs between sampling phases of fluorescence emitted along the first direction; each sampling phase in the first and second sets of sampling phases includes a voltage measurement of light from multiple light pulses emitted from a light source; and obtain multiple voltage measurements from each light pulse, wherein the multiple voltage measurements from each light pulse... The measurements include a first set of voltage measurements when the light source is on, and a second set of voltage measurements when the light source is off; the average of the first set of voltage measurements is calculated; the average of the second set of voltage measurements is calculated; the light pulse difference for each light pulse is determined by subtracting the average of the second set of voltage measurements from the average of the first set of voltage measurements; the voltage value for each sampling phase is determined by calculating the average of the light pulse differences for multiple light pulses in each sampling phase; the first polarization value for the first sampling phase is determined by calculating the average of the voltage values ​​in the first sampling phase; the second polarization value for the second sampling phase is determined by calculating the average of the voltage values ​​in the second sampling phase; and the concentration value of IgG is determined by subtracting the second polarization value from the first polarization value and then dividing by the sum of the first and second polarization values.

[0009] On the other hand, a system for measuring antibody concentration in a sample is disclosed, the system comprising: a light source; an excitation polarizer that polarizes light emitted from the light source along a first direction; a container configured to receive light polarized along the first direction; a polarizer holder that holds a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within the container to pass through in the first direction, and the second detection polarizer restricting fluorescence emitted from within the container to pass through in a second direction perpendicular to the first direction; and a detector for...The measurement includes fluorescence emitted along a first direction and fluorescence emitted along a second direction; and a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: rotate a polarizer holder about a rotation axis in alternating directions, such that a first detection polarizer and a second detection polarizer are alternately placed in the optical path of fluorescence emitted from within a container. Specification 2 / 23 pages 6 CN 121399448 A

[0010] On the other hand, a polarizer holder for a cell analysis system is disclosed, comprising: a first side surface; a second side surface perpendicular to the first side surface; a curved surface connecting the first and second side surfaces; a first detection polarizer coupled to the first side surface, the first detection polarizer allowing emitted fluorescence to pass in a first direction; and a second detection polarizer coupled to the second side surface, the second detection polarizer restricting emitted fluorescence to pass in a second direction; the polarizer holder is configured to rotate about a rotation axis, such that the first and second detection polarizers are alternately placed in the optical path of light.

[0011] On the other hand, a method for measuring the concentration of an antibody in a sample by fluorescence polarization is provided, the method comprising: positioning a polarizer holder in a first position such that a first detection polarizer is placed in the optical path of fluorescence emitted from inside a container, the container containing an antibody sample mixed with a fluorescence polarizing reagent; detecting fluorescence emitted from inside the container after light passes through the first detection polarizer, the first detection polarizer restricting the fluorescence emitted from inside the container to pass through in a first direction; rotating the polarizer holder about a rotation axis to position the polarizer holder in a second position such that a second detection polarizer is placed in the optical path of fluorescence emitted from inside the container, the second detection polarizer restricting the fluorescence emitted from inside the container to pass through in a second direction; and detecting fluorescence emitted from inside the container after light passes through the second detection polarizer.

[0012] On the other hand, a module for a cell analysis system is provided, the module comprising: a base having an internal volume for receiving an internal reservoir; a main cover connected to the base, the main cover including a port providing access to the internal reservoir; a secondary cover attached to the main cover, the secondary cover being configured to seal the port on the main cover; a motor attached to the secondary cover; and a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to: operate the motor to open the secondary cover, thereby allowing a probe to be inserted through the port on the main cover to reach the bottom of the internal reservoir; and when the probe is removed, operate the motor to close the secondary cover, thereby sealing the port on the main cover.

[0013] Various additional aspects will be set forth in the following description. These aspects may relate to individual features and combinations of features. It should be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only, and are not limiting.The embodiments disclosed herein are based on a broad inventive concept. Brief Description of the Drawings

[0014] The following drawings, which form part of this application, are illustrations of the described techniques and are not intended to limit the scope of this disclosure in any way.

[0015] FIG1 is an isometric view of an example of a cell analysis system for analyzing the cell health of multiple cell samples.

[0016] FIG2 is another isometric view of the cell analysis system of FIG1 with the top cover removed from the housing of the cell analysis system.

[0017] FIG3 is a top view of a working platform supported inside the housing of the cell analysis system of FIG1. ​​

[0018] FIG4 schematically shows an example of a titer module supported on the working platform of FIG3.

[0019] FIG5 graphically shows an example of a graph illustrating the error in calculating antibody concentration caused by detected light intensity drift in the titer module of FIG4.

[0020] FIG6 graphically shows a graph illustrating the simulated fluorescence polarization measurement error generated by the measurement sequence shown in the curve of FIG5.

[0021] FIG7 schematically shows an example of a method for measuring antibody concentration in a cell sample that can be performed by the titer module of FIG4.

[0022] Figure 8 schematically illustrates an example of a method for performing a measurement cycle in the operation of the method of Figure 7.

[0023] Figure 9 schematically illustrates an example of a measurement cycle performed according to the method of Figure 8.

[0024] Figure 10 graphically illustrates an example of a graph showing how the method of Figure 7 reduces the error in calculating the antibody concentration in a cell sample.

[0025] Figure 11 graphically illustrates a graph showing the reduction in fluorescence polarization measurement error caused by the measurement cycles of Figures 8 through 10.

[0026] Figure 12 schematically illustrates a method for measuring the sampling phase in the measurement cycles of Figures 8 through 10.

[0027] Figure 13 schematically illustrates an example of a controller for the cell analysis system of Figure 1, which can be used to implement various aspects of the titer module of Figure 4.

[0028] Figure 14 is an isometric view of an example of a titer module that can be used in the cell analysis system of Figure 1.

[0029] FIG15 is an isometric view of the titration module of FIG14, wherein a portion of the housing has been removed, exposing the internal components of the titration module.

[0030] FIG16 is an isometric cross-sectional view taken along the horizontal axis of the titration module of FIG14.

[0031] FIG17 is an isometric view of the titration module of FIG14, wherein the inner cover has been removed, exposing the optical components of the titration module.

[0032] FIG18 is a right-side view of the optical components of the titration module of FIG14.

[0033] FIG19 is a left-side view of the optical components of the titration module of FIG14.

[0034] FIG20 is an isometric cross-sectional view taken along the vertical axis of the titration module of FIG14.

[0035] Figure 21 is a right-side cross-sectional view of the titration module of Figure 14.

[0036] Figure 22 is a left-side cross-sectional view of the titration module of Figure 14.

[0037] Figure 23 is an isometric view of the polarizer holder, optical plate, and detector of the titration module of Figure 14.

[0038] Figure 24 is another isometric view of the polarizer holder, optical plate, and detector of the titration module of Figure 14.

[0039] Figure 25 is an exploded view of the polarizer holder of the titration module of Figure 14.

[0040] Figure 26 is an exploded view of the polarizer holder of the titration module of Figure 14.

[0041] Figure 27 is a top view of the polarizer holder of the titration module of Figure 14.

[0042] Figure 28 is an isometric view of the optical plate and motor of the titration module of Figure 14, with the polarizer holder removed therefrom.

[0043] Figure 29 is a top view of the optical plate of Figure 26.

[0044] FIG30 is an isometric view of the liquid storage module of the cell analysis system of FIG1. ​​

[0045] FIG31 is another isometric view of the liquid storage module of FIG30, showing the lid of the liquid storage module in the open position.

[0046] FIG32 is another isometric view of the liquid storage module of FIG30, showing the internal reservoir removed from the interior of the liquid storage module.

[0047] FIG33 is another isometric view of the liquid storage module of FIG30, showing the automatic lid of the liquid storage module in the open position.

[0048] FIG34 is another isometric view of the liquid storage module of FIG30, showing the lid of the liquid storage module in the open position.

[0049] FIG35 is an isometric view of the secondary lid of the liquid storage module of FIG30.

[0050] FIG36 is a front view of the secondary lid of FIG35.

[0051] FIG37 is a side view of the secondary lid of FIG35.

[0052] FIG38 is a bottom view of the secondary lid of FIG35.

[0053] Figure 39 is an isometric view of the interior of the cell analysis system of Figure 1, showing the dispensing system positioning the probe near the liquid storage module of Figure 30 (page 4 / 23 of specification, CN 121399448 A).

[0054] Figure 40 is an isometric view of the interior of the cell analysis system of Figure 1, showing the dispensing system inserting the probe into the internal reservoir of the liquid storage module of Figure 30.

[0055] Figure 41 shows an example of an alternative implementation of the polarizer holder of the titer module of Figure 14. Detailed Description

[0056] Figure 1 is an isometric view of an example of a cell analysis system 100 for analyzing cell health of multiple cell samples. For a single cell sample, the cell analysis system 100 can measure cell count, cell viability, antibody concentration (e.g., protein titer), and other cellular characteristics with minimal interaction with the system user.

[0057] The cell analysis system 100 includes connectivity with automated bioreactors and other systems and devices.The connectivity of automated bioreactors and other systems and devices may include electrical connectivity for sharing data, and may also include physical connectivity for liquid handling purposes. Cell analysis system 100 automates sample preparation and minimizes the sample volume required for measuring cell characteristics. Cell analysis system 100 provides remote access to data including the measured cell characteristics, supports multiple users, and is compatible with various information technology (IT) architectures.

[0058] As shown in FIG1, cell analysis system 100 includes a housing 102 supporting a work platform 300. Housing 102 includes a top cover 104 that can support a dispensing system 106, which will be described in more detail with reference to FIGS. 2 and 3.

[0059] FIG2 is another isometric view of cell analysis system 100, wherein the top cover 104 is removed from housing 102, thereby exposing the dispensing system 106. FIG3 is a top view of the work platform 300 supported inside housing 102. Referring now to Figures 2 and 3, the work platform 300 supports one or more tube racks that hold multiple containers containing cell samples and various types of reagents. In some cases, at least some of the containers are empty. The dispensing system 106 includes a probe 108, which is movably mounted within a housing 102 in the space above the work platform 300. The dispensing system 106 is an example of an automated pipetting robot system.

[0060] The probe 108 is mounted for movement along three mutually perpendicular axes (e.g., the X, Y, and Z axes of a three-dimensional Cartesian coordinate system). This three-dimensional movement allows the distal end of the probe 108 to approach any container held on the work platform 300 within the housing 102, through which liquid aspiration and dispensing are performed.

[0061] The proximal end of the probe 108 is fluidly connected to a bidirectional pump with a movable actuator that controls the pump's mode. For example, the first mode may include aspiration and the second mode may include dispensing, and a movable actuator controls the switching between the first and second modes, as well as the rate of aspiration and dispensing of liquid in the first and second modes. As an example, the bidirectional pump may include a syringe pump. The movement of the probe 108 and the movable actuator is controlled by one or more stepper motors, which operate under the control of a programmable controller 1300.

[0062] As shown in FIG2, the housing 102 is sized to have a width W, a depth D, and a height H. As an illustrative example, the width W is about 24 inches to about 36 inches, the depth D is about 24 inches to about 36 inches, and the height H is about 24 inches to about 36 inches. In some examples, the housing 102 is sized to have a cubic shape.

[0063] As shown in FIG3, the work platform 300 supports the sample transfer module 302, one or more mixing plates 304, and cell bridges.The module includes a cell health module 306, one or more tip holders 308, one or more titration plates 310, a tip waste bin 312, a liquid storage module 314, a sample input 316 (e.g., tube trays and well plates), a metabolite module 318, and a titration module 400. The cell health module 306 measures cell health, including cell count and cell viability. The titration module 400 measures protein titers and will now be described in more detail with reference to FIG4.

[0064] FIG4 schematically shows an example of a titration module 400 supported on a work platform 300. The titration module 400 includes optical components for measuring the concentration of antibodies present in a cell sample. For example, the titration module 400 measures the concentration of immunoglobulin G (IgG) present in a cell sample.

[0065] More specifically, the titration module 400 calculates fluorescence polarization measurements to measure the IgG protein concentration in the cell sample. Fluorescence polarization involves mixing a cell sample with a fluorescent polarizing reagent and subsequently measuring the fluorescence polarization to determine the concentration of IgG protein in the cell sample. While the titer module 400 is described herein with reference to the measurement of IgG concentration, the titer module 400 and the measurement technique described herein can be used to measure the concentrations of other types of proteins and antibodies.

[0066] As shown in FIG4, the titer module 400 includes a light source 402 that emits light 404 into a container 412 containing a cell solution mixed with the fluorescent polarizing reagent. The fluorescent polarizing reagent binds to target antibodies (e.g., IgG) produced by the cells. As discussed above, the dispensing system 106 of the cell analysis system 100 is automated to mix the fluorescent polarizing reagent with the cell sample in the container 412, so that the user of the system does not need to manually mix the solution in the container 412. In some examples, the container 412 is a cuvette, test tube, etc.

[0067] The light source 402 emits light 404 without polarization, such that the light 404 is unpolarized light. In some examples, the light source 402 is a light-emitting diode (LED). In some examples, the titration module 400 includes a focusing lens 406 and a spectral filter 408 that focus and filter light 404 emitted from the light source 402, respectively.

[0068] Light 404 passes through an excitation polarizer 410, which polarizes light 404 along a first direction 411. In some examples, the first direction 411 is a linear direction. In the example shown in FIG. 4, the first direction 411 is vertically linear. In an alternative example, the first direction 411 is horizontally linear. Other polarization directions are possible.

[0069] Once the light 404 is polarized along the first direction 411, it is absorbed by the cell solution in container 412 mixed with a fluorescent polarizing agent. This causes the solution in container 412 to emit polarized fluorescence 416.

[0070] Polarized fluorescence 416 can be focused by passing through lens 414 after being emitted from the solution in container 412. The polarized fluorescence 416 then alternately passes through a first detection polarizer 420 and a second detection polarizer 422. The first detection polarizer 420 and the second detection polarizer 422 are mounted to a polarizer holder 418, which is controlled by controller 1300 to alternately position the first detection polarizer 420 and the second detection polarizer 422 in the path of the polarized fluorescence 416.

[0071] The first detection polarizer 420 is polarized in a direction parallel to the direction of the excitation polarizer 410 (i.e., in a first direction 411). The first detection polarizer 420 confines the polarized fluorescence 416 to pass through in the first direction 411.

[0072] The second detection polarizer 422 is polarized in a second direction 413 perpendicular to the first direction 411 of the excitation polarizer 410. The second detection polarizer 422 allows the polarized fluorescence 416 to pass through in the second direction 413. Subsequently, polarized fluorescence 416 passes through spectral filter 424 before being detected by detector 426. In some examples, detector 426 is a photomultiplier tube (PMT). Detector 426 converts polarized fluorescence 416 into a voltage value, which is input into Equation 1 to determine the antibody concentration produced by the cell sample.

[0073]

[0074] Wherein IPAR is the voltage of parallel fluorescence detected from the solution in container 412, IPER is the voltage of vertical fluorescence detected from the solution in container 412, and FP is the fluorescence polarization in millipoles (mP). As an example, the voltages of IPAR and IPER detected by detector 426 can range from about 0.02 volts to about 5.0 volts.

[0075] The measured fluorescence polarization degree (FP) is related to the antibody concentration in the cell sample. For example, a higher detected FP is associated with a higher concentration of antibody, while a lower detected FP is associated with a lower concentration of antibody. This is because a higher concentration of antibody will increase the binding to the fluorescent polarizing reagent, resulting in a larger voltage for parallel fluorescence and a smaller voltage for vertical fluorescence, which leads to a larger detected FP.

[0076] The technical challenge of Equation 1 is that any error in measuring IPAR (which does not exist in IPER) may lead to calculation errors in fluorescence polarization specification page 6 / 23 10 CN 121399448 A (FP). For example, FP calculation depends on the measured ratio IPER / IPAR. Any changes that proportionally affect IPAR and IPER will be canceled out. However, errors such as signal intensity changes during the measurement of parallel fluorescence (IPAR) and vertical fluorescence (IPER) and the offset errors affecting both vertical fluorescence (IPER) and parallel fluorescence (IPAR) will not be canceled out and will cause significant errors in FP calculation, and thus errors in calculating antibody concentration in cell samples.

[0077] Sources of signal intensity variation may include, but are not limited to, intensity fluctuations and drift of light source 402, electrical response, dye bleaching, and non-uniform fluid and diffusion in container 412. Additional sources of error are also possible. Sources of offset noise may include, but are not limited to, circuit noise and ambient light.

[0078] Figure 5 graphically illustrates an example of a graph 500 showing the error in FP calculation and the incorrect calculation of antibody concentration in the cell sample caused by light intensity drift detected in titration module 400. Graph 500 includes light intensity (Y-axis) measured by detector 426 over time (X-axis). In this example, the light intensity is measured without fluorescent polarizing reagents and any cell solution in container 412, such that the light intensity drift is not caused by the solution.

[0079] In this example, the light intensity of light source 402 is expected to have a constant value of 3.5 V. Therefore, since there is no cell sample containing antibodies in container 412, the ratio between IPER fluorescence and IPAR fluorescence should be 1:1 (e.g., 3.5 V: 3.5 V).

[0080] However, the actual light intensity shown in graph 500 exhibits voltage drift, which leads to an error in the ratio between IPER fluorescence and IPAR fluorescence. In this example, a measurement cycle is performed in which IPAR fluorescence is measured in a first phase (i.e., using the second detection polarizer 422) and subsequently in a second phase (i.e., using the first detection polarizer 420). Each of the IPER and IPAR fluorescence phases includes two measurements. For example, IPER fluorescence includes a first measurement of 3.60 V and a second measurement of 3.62 V, and IPAR fluorescence includes a third measurement of 3.64 V and a fourth measurement of 3.66 V. IPER fluorescence is calculated as the average of the first and second measurements (i.e., 3.61 V), and IPAR fluorescence is calculated as the average of the third and fourth measurements (i.e., 3.65 V). This results in a ratio of 3.61:3.65 between IPER and IPAR fluorescence, which is an error of approximately 1% caused by the drift of light source 402.

[0081] Figure 6 graphically illustrates a graph 600 showing the simulated fluorescence polarization measurement error generated by the measurement sequence of Figure 5. As shown in Figure 6, the measurement error is significantly larger for lower mP values, such as mP values ​​less than 200 units. Therefore, lower concentrations of antibody in the sample contained in the container are particularly sensitive to signal intensity variations and noise in the titration module 400.

[0082] In some cases, the measurement error of the ratio between IPER fluorescence and IPAR fluorescence due to signal intensity variations and noise has been reduced by implementing complex circuit designs and temperature-controlled light sources and detectors in the titration module. However, such solutions are expensive, unreliable, may increase the size of the titration module, and slow down the warm-up time of the device.

[0083] Figure 7 schematically illustrates an example of a method 700 for measuring antibody concentration in a cell sample. Method 700 uses the optical components of titer module 400 (as shown in Figure 4) to calculate the antibody concentration in the cell sample. In some examples, method 700 is performed to measure the concentration of immunoglobulin G (IgG) in a solution containing a fluorescent polarizing reagent, by mixing the solution containing the fluorescent polarizing reagent in container 412 using dispensing system 106 of cell analysis system 100.

[0084] Method 700 reduces errors from signal intensity variations and noise without requiring changes to the optical components and hardware of titer module 400. In fact, method 700 is a software solution that improves the functionality of titer module 400 by reducing these types of errors to improve the accuracy of antibody concentration calculation.

[0085] Method 700 includes an operation 702 for performing a measurement cycle. Operation 702 includes acquiring a first set of measurements of fluorescence emitted from the solution along a first direction 411 using the optical components of titer module 400, and acquiring a second set of measurements of fluorescence emitted from the solution along a second direction 413. During the measurement cycle, the first set of measurements and the second set of measurements alternate.

[0086] FIG8 schematically illustrates an example of a method 800 performing a measurement cycle in operation 702 of method 700. FIG9 schematically illustrates an example of a measurement cycle 900 performed according to method 800.

[0087] Referring now to FIG8 and FIG9, method 800 includes step 802 of measuring a first sampling phase A of fluorescence emitted from container 412 along a first direction 411 before the midpoint 902 of measurement cycle 900. Next, method 800 includes step 804 of measuring a second sampling phase B of fluorescence emitted from container 412 along a second direction 413 before the midpoint 902 of measurement cycle 900.

[0088] As further illustrated in Figures 8 and 9, method 800 includes step 806 of measuring a third sampling phase C of fluorescence emitted from container 412 along a second direction 413 after the midpoint 902 of measurement period 900. Next, method 800 includes step 808 of measuring a fourth sampling phase D of fluorescence emitted from container 412 along a first direction 411 after the midpoint 902 of measurement period 900.

[0089] Measurement period 900 includes at least one sampling phase along the first direction 411 and at least one sampling phase along the second direction 413 before the midpoint. Measurement period includes at least one sampling phase along the first direction 411 and at least one sampling phase along the second direction 413 after the midpoint of measurement period.

[0090] During measurement period 900, polarizer holder 418 is controlled by controller 1300 to, at transition point 904, from along...A first detection polarizer 420, which is directionally polarized parallel to the direction of the excitation polarizer 410, is switched to a second detection polarizer 422, which is directionally polarized perpendicular to the direction of the excitation polarizer 410. A transition point 904 occurs between the first sampling phase A and the second sampling phase B. Furthermore, during the measurement period 900, the polarizer holder 418 is controlled by the controller 1300 to switch at a transition point 906 from the second detection polarizer 422 (which is directionally polarized perpendicular to the direction of the excitation polarizer 410) to the first detection polarizer 420 (which is directionally polarized parallel to the direction of the excitation polarizer 410). A transition point 906 occurs between the third sampling phase C and the fourth sampling phase D.

[0091] As further shown in FIG9, each of the sampling phases A to D includes a voltage measurement of the fluorescence from a plurality of light pulses 908 emitted from the light source 402. For each of the light pulses 908, the detector 426 obtains a plurality of voltage measurements 910. For example, the plurality of voltage measurements 910 from each of the light pulses 908 include a first set of voltage measurements 910a when the light source 402 is on and a second set of voltage measurements 910b when the light source 402 is off. Steps 802 to 808 for measuring the first sampling phase A, the second sampling phase B, the third sampling phase C, and the fourth sampling phase D, respectively, may each include performing the additional operations shown in FIG12.

[0092] FIG12 schematically illustrates a method 1200 for measuring sampling phases performed in the measurement cycle of FIG8 to FIG10. Referring now to FIG9 and FIG12, method 1200 includes an operation 1202 of calculating the average value of the first set of voltage measurements 910a when the light source 402 is on. Next, method 1200 includes an operation 1204 of calculating the average value of the second set of voltage measurements 910b when the light source 402 is off. Next, method 1200 includes an operation 1206 of determining the voltage value of each of the light pulses 908 in the sampling phase by subtracting the average value of the second set of voltage measurements 910b from the average value of the first set of voltage measurements 910a. Next, method 1200 may include an operation 1208 of determining the voltage value of the sampling phase by calculating the average of the voltage values ​​of the plurality of light pulses 908 in the sampling phase.

[0093] Referring back to FIG7, method 700 includes an operation 704 of calculating a first average value of a first sampling phase A and a fourth sampling phase D. As shown in FIG9, the first sampling phase A includes fluorescence emitted along a first direction 411 before the midpoint 902 of the measurement period 900. The fourth sampling phase D includes fluorescence emitted along the first direction 411 after the midpoint 902 of the measurement period 900.

[0094] Next, method 700 includes an operation 706 of calculating a second average value of a second sampling phase B and a third sampling phase C. As described above, the second sampling phase B includes fluorescence emitted from the container along a second direction before the midpoint 902 of the measurement period 900.Fluorescence towards 413. The third sampling phase C includes fluorescence emitted from the container along the second direction 413 after the midpoint 902 of the measurement cycle 900. Specification 8 / 23 pages 12 CN 121399448 A

[0095] Method 700 includes an operation 708 to determine the antibody concentration based on the ratio of a first average and a second average. For example, the first average is the voltage of parallel fluorescence (IPAR), the second average is the voltage of vertical fluorescence (IPER), and the antibody concentration is calculated using Formula 1.

[0096] Figure 10 graphically illustrates an example of a graph 1000 showing how method 700 eliminates errors in the calculation of polarization measurement units (mP) used to determine the antibody concentration in a cell sample. Graph 1000 includes light intensity (Y-axis) measured by detector 426 over time (X-axis). Similar to the example graph in Figure 5, graph 1000 shows that the light source 402 exhibits a drift relative to the expected value of 3.5 V. The light intensity measurements were performed without fluorescent polarizing reagents and any cell solution in container 412, therefore the light intensity drift was not caused by the solution.

[0097] As shown in FIG10, a measurement cycle was performed, with the first sampling phase A (3.55 V) for fluorescence emitted in the first direction before the midpoint M, the second sampling phase B (3.59 V) for fluorescence emitted in the second direction before the midpoint M, the third sampling phase C (3.61 V) for fluorescence emitted in the second direction after the midpoint M, and the fourth sampling phase D (3.65 V) for fluorescence emitted in the first direction after the midpoint M. The first average of the first sampling phase A and the fourth sampling phase D was calculated to be 3.6 V. The second average of the second sampling phase B and the third sampling phase C was calculated to be 3.6 V. This resulted in a ratio of 3.6:3.6 between IPER fluorescence and IPAR fluorescence, which eliminated the error caused by the drift of light source 402 (see curve 500 in FIG5 for comparison). Therefore, the measurement cycle performed in method 800 reduces the error when measuring antibody concentration in cell samples using fluorescence polarization, and even eliminates the error in some cases.

[0098] Figure 11 graphically illustrates a graph 1100 showing the reduction in simulated fluorescence polarization measurement error caused by the measurement cycles shown in Figures 8 through 10. Referring now to Figures 6 and 11, graph 1100 shows that for lower mP values ​​(e.g., mP values ​​less than 200 units), the measurement error is significantly smaller than the measurement error shown in graph 600 when performing conventional measurement sequences. Graph 1100 shows a significant reduction in sensitivity to signal intensity variations and noise in the titer module 400 for lower antibody concentrations.

[0099] Figure 13 schematically illustrates an example of a controller 1300 of a cell analysis system 100, which can be used to implement the aspects described herein, including the features of the titer module 400. As shown in Figure 13, the controller 1300 includes one or more processing devices 1302, a memory storage device 1304, and a system bus 1306 coupling the memory storage device 1304 to one or more processing devices 1302. One or more processing devices 1302 may include a central processing unit (CPU). In some cases, one or more processing devices 1302 are part of a processing circuitry system having memory for storing instructions that, when executed by the processing circuitry system, cause the processing circuitry system to perform the various aspects, features, and functions described herein.

[0100] As shown in Figure 13, the memory storage device 1304 may include a random access memory (“RAM”) 1308 and a read-only memory (“ROM”) 1310. Basic input and output logic having basic routines that facilitate the transfer of information between elements within the controller 1300 (e.g., during startup) may be stored in the ROM 1310.

[0101] The controller 1300 may also include a mass storage device 1312, which may include an operating system 1314 and store software instructions and data 1316. The mass storage device 1312 is connected to the processing device 1302 via a system bus 1306. The mass storage device 1312 and the associated computer-readable data storage medium provide the controller 1300 with non-volatile, non-transitory storage.

[0102] Although the description of the computer-readable data storage medium contained herein refers to the mass storage device 1312, those skilled in the art will understand that the computer-readable data storage medium can be any available non-transitory physical device or article of manufacture from which the controller 1300 can read data and / or instructions. The computer-readable storage medium may consist entirely of non-transitory media. The mass storage device 1312 is an example of a computer-readable storage device. Specification page 9 / 23, CN 121399448 A

[0103] Computer-readable data storage media include volatile and non-volatile, removable and non-removable media implemented in any method or technology for storing information such as computer-readable software instructions, data structures, program modules or other data. Example types of computer-readable data storage media include, but are not limited to, RAM, ROM, EPROM, EEPROM, flash memory or other solid-state memory technologies, or any other medium that can be used to store information and can be accessed by a device.

[0104] Controller 1300 can operate in a networked environment using a logical connection to other devices via network 1320.The controller 1300 is connected to the network 1320 via a network interface unit 1318 connected to the system bus 1306. The network interface unit 1318 can also connect to additional types of communication networks and devices, including via Bluetooth, Wi-Fi, and cellular telecommunications networks (including 4G and 5G networks). The network interface unit 1318 can connect the controller 1300 to additional networks, systems, and devices. The controller 1300 also includes an input / output unit 1322 for receiving and processing inputs and outputs from peripheral devices.

[0105] The mass storage device 1312 and RAM 1308 can store software instructions and data. The software instructions may include an operating system 1314 suitable for controlling the operation of the cell analysis system 100. The mass storage device 1312 and / or RAM 1308 may also store software instructions and data 1216, which, when executed by the processing device 1302, provide the functionality of the cell analysis system 100 discussed herein.

[0106] FIG14 is an isometric view of an example of a titer module 400 that can be used in the cell analysis system 100 (see also FIG3 and FIG4, for example). FIG15 is an isometric view of the titer module 400 with a portion of the housing 401 removed, exposing the internal components of the titer module. FIG16 is an isometric cross-sectional view taken along the horizontal axis of the titer module 400. Referring now to FIGS. 14 through 16, the internal components of the titer module 400 include an excitation assembly 403 and a detection assembly 405.

[0107] The excitation assembly 403 includes a first optical cover 407 that at least partially houses a light source 402 and an excitation polarizer 410. The detection assembly 405 includes a second optical cover 409 that at least partially houses a detector 426, a polarizer holder 418, a first detection polarizer 420, and a second detection polarizer 422.

[0108] As will be described in more detail, the polarizer holder 418 is operated by the controller 1300 to precisely align the detection polarization axes of the first detection polarizer 420 and the second detection polarizer 422 to be parallel and perpendicular to the excitation polarization axis of the excitation polarizer 410, respectively. This arrangement is generally more precise than an arrangement with rotating polarizer axes, which can generally result in greater variability of the polarizer in the optical path. The polarizer holder 418 addresses the technical problem of repeatably and precisely switching between the first detection polarizer 420 and the second detection polarizer 422 on the detection leg of the optical path in the titration module 400.

[0109] FIG17 is an isometric view of the titration module 400, wherein the first optical cover 407 and the second optical cover 409 are removed, exposing the optical components of the titration module. FIG18 is a right-side view of the optical components of the titration module 400. FIG19 is a left-side view of the optical components of the titration module 400. The optical components of the titration module 400 shown in FIG17 to FIG19 are substantially similar to those in FIG18.The optical components are schematically shown in Figure 4. For example, the titration module 400 includes a light source 402 that emits light toward a container 412 containing a cell culture of IgG protein mixed with a fluorescent polarizing agent. In some examples, the container 412 is a cuvette, test tube, etc.

[0110] The light source 402 emits light unpolarized, such that the light is unpolarized. In some examples, the light source 402 is a light-emitting diode (LED). In some examples, the titration module 400 includes an optical retainer 460, which includes a focusing lens 406 and a spectral filter 408 that focus and filter the light emitted from the light source 402, respectively.

[0111] The light passes through an excitation polarizer 410 that polarizes the light along a first direction 411 (see Figure 4). Once the light is polarized along the first direction 411, it is absorbed by the cell solution in the container 412 mixed with the fluorescent polarizing agent. This causes the solution in the container 412 to emit polarized fluorescence.

[0112] After being emitted from container 412, the polarized fluorescence passes through lens 414 and aperture 415. The light then passes through either a first detection polarizer 420 or a second detection polarizer 422 mounted to a holder 418. As will be described in more detail, the polarizer holder 418 is powered by a motor 428 to rotate, which causes the first detection polarizer 420 and the second detection polarizer 422 to be alternately positioned in the path of the polarized fluorescence emitted from the solution in container 412.

[0113] As described above, the first detection polarizer 420 is polarized in a direction parallel to the direction of the excitation polarizer 410, while the second detection polarizer 422 is polarized in a direction perpendicular to the direction of the excitation polarizer 410.

[0114] The polarized fluorescence then passes through a spectral filter 424 before reaching detector 426. In some examples, detector 426 is a photomultiplier tube (PMT). As described above, detector 426 converts polarized fluorescence into a voltage value to be input into Equation 1 to determine the antibody concentration produced by the cell culture.

[0115] FIG20 is an isometric cross-sectional view taken along the vertical axis of titration module 400. FIG21 is a right-side cross-sectional view of titration module 400. FIG22 is a left-side cross-sectional view of titration module 400. Referring now to FIGS. 20 to 22, polarizer holder 418 is connected to drive shaft 434 of motor 428. In some examples, motor 428 is a stepper motor (also called a stepper motor or stepper motor) or a similar type of electric motor. In other examples, motor 428 is a solenoid. Motor 428 is mounted to the underside of optical plate 419 supporting the optical components of titration module 400.

[0116] Controller 1300 controls motor 428 to cause polarizer holder 418 to rotate clockwise around rotation axis R-R in the direction D1 andThe first detection polarizer 420 and the second detection polarizer 422 are rotated counterclockwise D2 (see Figure 27) so that they are alternately positioned in the optical path of the polarized fluorescence emitted from the solution in the container 412 before the light reaches the detector 426. In some examples, the controller 1300 controls the motor 428 to rotate the polarizer holder 418 about the rotation axis R-R clockwise D1 by about 90 degrees and counterclockwise D2 by about 90 degrees.

[0117] In an alternative embodiment, Figure 41 shows an example of a polarizer holder 418a, which includes a swivel mount 462 and an electromagnetic driver 464 that can be used to position the detection polarizer 466 in the path of the fluorescence emitted from the container 412. In some examples, the electromagnetic driver 464 drives the swivel mount 462 to rotate the detection polarizer 466 about an axis parallel to the beam entering the detection polarizer. In this alternative embodiment, the electromagnetic actuator 464 rotates the detection polarizer 466 to a first position, such that the detection polarizer 466 has a polarity parallel to the polarity of the excitation polarizer 410, and rotates the detection polarizer 466 to a second position, such that the detection polarizer 466 has a polarity perpendicular to the polarity of the excitation polarizer 410. In this example, the electromagnetic actuator 464 can rotate the detection polarizer approximately 90 degrees between the first and second positions. In this example, the electromagnetic actuator 464 includes a magnet 468 positioned adjacent to the swivel mount 462, and the magnet 468 drives the swivel mount 462 to rotate the detection polarizer 466 to a desired rotational position. In some examples, the magnet 468 surrounds or at least partially surrounds the swivel mount 462 and the detection polarizer 466. The electromagnetic actuator 464 can provide higher accuracy than rack and pinion mechanisms used for positioning the detection polarizer.

[0118] Figures 23 and 24 are isometric views of the polarizer holder 418, optical plate 419, and detector 426 of the titration module 400. Figures 25 and 26 are exploded views of the polarizer holder 418. Figure 27 is a top view of the polarizer holder 418. Referring now to Figures 23 through 27, the polarizer holder 418 has a turret-like arrangement. The polarizer holder 418 includes a first side surface 430 and a second side surface 432. In some examples, the first side surface 430 and the second side surface 432 are perpendicular to each other. The polarizer holder 418 also includes a curved surface 431 connecting the first side surface 430 and the second side surface 432. The exterior of the polarizer holder 418 includes the first side surface 430, the second side surface 432, and the curved surface 431.

[0119] A first detection polarizer 420 is mounted to a first side surface 430 of the polarizer holder 418, and a second detection polarizer 422 is mounted to a second side surface 432 of the polarizer holder 418. For example, the first side surface 430 includes a recess 435, the second detection polarizer 422 is mounted to a second side surface 432 of the polarizer holder 418.A detection polarizer 420 is placed in a recess 435. Similarly, a second side surface 432 includes a recess 435 in which a second detection polarizer 422 is placed. As shown in FIG27, the first detection polarizer 420 and the second detection polarizer 422 protrude beyond the corresponding first side surface 430 and second side surface 432 when placed inside the recess 435.

[0120] As shown in FIGS. 25 to 27, a biasing element 436 is used to hold the first detection polarizer 420 and the second detection polarizer 422 respectively within the recesses 435 of the first side surface 430 and the second side surface 432. Fasteners 438, such as screws, can be screwed through holes 440 on the bias element 436 into holes 441 on the first side surface 430 and the second side surface 432 to attach the bias element 436 to the first side surface 430 and the second side surface 432, and thereby secure the first detection polarizer 420 and the second detection polarizer 422 in the recess 435. In some examples, the bias element 436 is a retaining spring.

[0121] The bias elements 436 are flexible such that they are configured to bend around the edges of the first detection polarizer 420 and the second detection polarizer 422 to secure the first detection polarizer 420 and the second detection polarizer 422 in place. In some examples, the bias elements 436 include cutouts 439 to increase their flexure. In some examples, the cutouts 439 are positioned toward the top and bottom of each bias element 436 to increase the flexure of the bias elements 436 around the top and bottom of the first detection polarizer 420 and the second detection polarizer 422.

[0122] The biasing element 436 includes an aperture 442 that allows polarized fluorescence emitted from the solution in container 412 to pass through the first detection polarizer 420 and the second detection polarizer 422. Recesses 435 each include an aperture 437 (see FIG. 25) that aligns with a corresponding aperture 444 (see FIG. 26) on the curved surface 431, thereby allowing polarized fluorescence to pass through the polarizer holder 418 and reach the detector 426.

[0123] As shown in FIG. 16, the curved surface 431 of the polarizer holder 418 is aligned with the curved wall 417 of the second optical cover 409 to minimize the space between the polarizer holder 418 and the curved wall 417. This helps mitigate stray light entering the first detection polarizer 420 and the second detection polarizer 422 and reaching the detector 426, which may interfere with the voltage value detected by the detector 426. Therefore, the curved surface 431 of the polarizer holder 418 can reduce noise in the voltage value detected by the detector 426.

[0124] FIG28 is an isometric view of the optical plate 419 and motor 428 of the titration module 400, wherein the polarizer holder is located from itsRemove. Figure 29 is a top view of the optical plate 419. Referring now to Figures 28 and 29, as discussed above, the polarizer holder 418 is directly fixed to the drive shaft of the motor 428, and the motor is mounted on the underside of the optical plate 419. The polarizer holder 418 includes a locating pin 446 on the underside of the holder. As the polarizer holder 418 rotates about the rotation axis R-R in clockwise direction D1 and counterclockwise direction D2 (see Figures 20 to 22 and Figure 27), the locating pin 446 slides within a groove 448 in the optical plate 419. As shown in Figures 28 and 29, the groove 448 bends along the rotation axis R-R.

[0125] The locating pin 446 contacts a first stop 450 and a second stop 451 in the groove 448 at the end of each rotational stroke in the clockwise direction D1 and counterclockwise direction D2. For example, when the polarizer holder 418 is rotated 90 degrees clockwise D1 to position the first detection polarizer 420 in the optical path of light, the positioning pin 446 contacts the first stop 450 at the distal end of the groove 448 to prevent further rotation of the polarizer holder 418. Similarly, when the polarizer holder 418 is rotated 90 degrees counterclockwise D2 to position the second detection polarizer 422 in the optical path of light, the positioning pin 446 contacts the second stop 451 at the opposite distal end of the groove 448 to prevent further rotation of the polarizer holder 418.

[0126] Each of the first stop 450 and the second stop 451 is associated with the position of the polarizer holder 418. For example, when the first detection polarizer 420 is positioned in the optical path of light, the first stop 450 is associated with the parallel polarizer position 454 (see FIG. 27). When the second detection polarizer 422 is positioned in the optical path of the light, the second stop 451 is associated with the vertical position 456.

[0127] As further shown in Figures 28 and 29, the optical plate 419 includes a magnet 452 positioned next to each of the first stop 450 and the second stop 451. The positioning pin 446 is made of a ferromagnetic material such as steel, such that when positioned near the first stop 450 at the end of a rotational stroke in the clockwise direction D1, the magnet 452 attracts the positioning pin 446, and when positioned near the second stop 451 at the end of a rotational stroke in the counterclockwise direction D2, the magnet 452 attracts the positioning pin 446.

[0128] The controller 1300 is programmed to shut off the motor 428 at the end of the rotational stroke in the clockwise direction D1 and the counterclockwise direction D2, causing the magnet 452 to firmly pull the positioning pin 446 upward against the first stop 450 and the second stop 451. The magnet 452 ensures a high degree of repeatability between the parallel polarizer position and the vertical polarizer position, and improves the overall accuracy and repeatability of the titration module 400.

[0129] The arrangement of the polarizer holder 418, rotating clockwise D1 and counterclockwise D2 to alternately place the first detection polarizer 420 and the second detection polarizer 422 in the optical path of light emitted from the solution in the container 412, reduces the space required for the polarizer holder 418 on the optical plate 419 of the titration module 400. Furthermore, the arrangement of the polarizer holder 418 reduces stray light passing through the first detection polarizer 420 and the second detection polarizer 422, and ensures that the alignment of the first detection polarizer 420 and the second detection polarizer 422 remains fixed relative to the excitation polarization axis of the excitation assembly 403 from the titration module 400.

[0130] FIG30 is an isometric view of the liquid storage module 314 of the cell analysis system 100 (see also FIG3). In FIG30, the liquid storage module 314 is shown in the closed position. The liquid storage module 314 can be used in combination with the titration module 400. For example, the liquid storage module 314 can contain liquids used by the titration module 400, such as diluents. The liquid storage module 314 is designed to prevent evaporation of the liquid diluent (especially liquid diluents sensitive to light and air exposure) while allowing efficient loading and pipetting of the liquid diluent. As an illustrative example, the liquid storage module 314 can limit evaporation to less than about 1.5% (volume) for an onboard storage lifetime of about 31 days within the cell analysis system 100.

[0131] Figures 31 and 32 are isometric views showing the liquid storage module 314 in the open position. Referring now to Figures 30-32, the liquid storage module 314 includes a base 3002 having an internal volume 3004 for accommodating an internal reservoir 3006. A user can access the internal reservoir 3006 by manually opening the main cover 3008 (e.g., to load liquid diluent into the internal reservoir 3006). The internal reservoir 3006 is reusable and replaceable. For example, the internal reservoir 3006 can be removed from the internal volume 3004, for example, for cleaning or replacement with another internal reservoir (e.g., a new or clean internal reservoir).

[0132] In some examples, the internal reservoir 3006 comprises a single well with a volume of approximately 40 mL. In some examples, the internal reservoir 3006 has a V-shaped bottom. Once the diluent has been loaded into the internal reservoir 3006, the user can manually close the main cap 3008. The main cap 3008 remains closed during operation of the cell analysis system 100.

[0133] The main cap 3008 can be manually pivoted about the hinge 3012 to move between a closed position (FIG. 30) and an open position (FIGs. 31 and 32). The liquid storage module 314 includes a gasket 3014 surrounding the internal volume 3004 that houses the internal reservoir 3006. The gasket 3014 may be made of ethylene propylene diene monomer (EPDM) rubber. When the main cover 3008 is in the closed position, the gasket3014 engages the inner surface 3016 of the main cover 3008 to seal around the internal reservoir 3006.

[0134] The main cover 3008 includes one or more first magnets 3018 that are magnetically attracted to one or more second magnets 3020 positioned on the base 3002. Once the main cover 3008 is pivoted about the hinge 3012 to the closed position, the attraction of the one or more first magnets 3018 to the one or more second magnets 3020 can keep the cover closed.

[0135] FIG33 is another isometric view of the liquid storage module 314 showing the main cover 3008 in the closed position. Referring now to FIG30 and FIG33, the liquid storage module 314 includes a secondary cover 3010 attached to the main cover 3008. The secondary cap 3010 is controlled by the controller 1300 to open and close as needed by the probe, so that the probe 108 of the dispensing system 106 (see FIG. 2) is close to the internal reservoir 3006. For example, the controller 1300 opens the secondary cap 3010 during pipetting and closes it after pipetting is complete, to seal the diluent within the internal reservoir 3006 and thus prevent the diluent from evaporating during use of the cell analysis system 100.

[0136] As shown in FIG. 33, when the secondary cap 3010 is opened, the port 3022 on the main cap 3008 is exposed. The probe 108 of the dispensing system 106 can be inserted through the port 3022 to access the internal reservoir 3006 without opening the main cap 3008. In some examples, the port 3022 is passed through a tapered pipette tip sized to 55 μL or 230 μL. This type of pipette is generally not compatible with solid diaphragm caps.

[0137] The secondary cap 3010 has a gasket 3024 to seal the port 3022 when the secondary cap 3010 is in the closed position to prevent evaporation of the diluent contained in the internal reservoir 3006. In some examples, the gasket 3024 is made of polyethylene (PE) foam.

[0138] FIG34 is another isometric view of the liquid storage module 314 showing the main cap 3008 in the open position. Referring now to FIG34, a magnet 3026 is mounted to the main cap 3008, which attracts the iron-containing element 3028 on the secondary cap 3010 to close the secondary cap 3010. For example, the hinge 3030 of the secondary cap 3010 may include the iron-containing element 3028 attracted to the magnet 3026 to close the secondary cap 3010. In an alternative example, magnet 3026 is included on sub-cover 3010, and ferro-containing element 3028 is included on main cover 3008. Magnet 3026 and ferro-containing element 3028 keep sub-cover 3010 closed.

[0139] Motor 3032 is mounted to the inner surface 3016 of main cover 3008. Sub-cover 3010 is connected to motor 3032. Motor 3032 can...This may include a stepper motor or a similar type of electric motor. Motor 3032 is controlled by controller 1300 to move the sub-cover 3010 between an open and closed state. Motor 3032 may utilize an integrated encoder to track the open and closed states of the sub-cover 3010.

[0140] FIG35 is an isometric view of the sub-cover 3010. FIG36 is a front view of the sub-cover 3010. FIG37 is a side view of the sub-cover 3010. FIG38 is a bottom view of the sub-cover 3010. Referring now to FIGS. 35 to 38, the sub-cover 3010 includes a top portion 3040 and a hinge 3042 extending from the top portion. When the sub-cover 3010 is in the closed state, the top portion 3040 covers the port 3022. The hinge 3042 is connected to the motor 3032. For example, the hinge 3042 includes a first hole 3044, which can be used to connect the sub-cover 3010 to the drive shaft of the motor 3032. The hinge 3042 may also include a second hole 3046, for example for attaching a ferrous element 3028 (e.g., a screw), which is attached to a magnet 3026 on the main cover 3008 to keep the secondary cover 3010 closed.

[0141] In some examples, the secondary cover 3010 includes a plunger 3048 extending from the bottom surface 3050 of the top 3040 such that the plunger is inserted into the interior of the port 3022 when the secondary cover 3010 is in the closed state. Additionally, a gasket 3024 may be attached around the plunger 3048 to seal the port 3022 and mitigate diluent evaporation.

[0142] FIG39 is an isometric view of the interior of the cell analysis system 100, showing the dispensing system 106 positioning the probe 108 behind the liquid storage module 324. FIG40 is an isometric view of the interior of the cell analysis system 100, showing the dispensing system 106 and the liquid storage module 324, with the secondary cover 3010 in the open state. The design of the liquid storage module 324 includes a low profile when the main cap 3008 is in the closed position, and a secondary cap 3010 that automatically opens and closes as needed. This design allows the dispensing system 106 to move the probe 108 freely above the work platform 300 inside the cell analysis system 100, so that the probe 108 can approach laboratory glassware placed behind the liquid storage module 324 without colliding with it. Furthermore, the design of the liquid storage module 324 allows the dispensing system 106 to fully extend the probe 108 (e.g., a 50 μL or 230 μL pipette) through the port 3022 to reach the bottom of the internal reservoir 3006 and draw diluent, while also mitigating diluent evaporation during use of the cell analysis system 100 by closing the secondary cap 3010 and sealing the port 3022 after diluent draw.

[0143] The various embodiments described above are provided by way of illustration only and should not be construed as being in any way...Limitations may be imposed. Various modifications may be made to the above embodiments without departing from the true spirit and scope of this disclosure.

[0144] Embodiments of this disclosure may be described with reference to the following numbered clauses, wherein preferred features are arranged in the clauses of CN 121399448 A from page 14 / 23 of the specification:

[0145] 1. A method for measuring the fluorescence polarization of an antibody concentration in a sample, the method comprising:

[0146] performing a measurement cycle, measuring a first set of sampling phases of fluorescence emitted along a first direction and a second set of sampling phases of fluorescence emitted along a second direction, at least one sampling phase of the fluorescence emitted along the second direction occurring between sampling phases of the fluorescence emitted along the first direction;

[0147] calculating a first value of the first set of sampling phases;

[0148] calculating a second value of the second set of sampling phases; and

[0149] determining the antibody concentration based on a function of the first value and the second value.

[0150] 2. According to the method of Clause 1, wherein a first sampling phase of the first set of sampling phases of the fluorescence emitted along the first direction occurs before the midpoint of the measurement period, and a second sampling phase of the first set of sampling phases of the fluorescence emitted along the first direction occurs after the midpoint of the measurement period.

[0151] 3. According to the method of Clause 1, wherein each sampling phase includes voltage measurements of fluorescence from a plurality of light pulses emitted from the light source.

[0152] 4. According to the method of Clause 3, further comprising:

[0153] obtaining a plurality of voltage measurements from each light pulse.

[0154] 5. According to the method of Clause 4, wherein the plurality of voltage measurements from each light pulse includes a first set of voltage measurements when the light source is turned on and a second set of voltage measurements when the light source is turned off.

[0155] 6. The method according to Clause 5 further includes:

[0156] calculating a first average value of the first set of voltage measurements;

[0157] calculating a second average value of the second set of voltage measurements; and

[0158] determining a light pulse difference for each light pulse by subtracting the second average value from the first average value.

[0159] 7. The method according to Clause 6 further includes:

[0160] determining an average light pulse difference for the sampled phase by calculating an average of the light pulse differences from the sampled phase.

[0161] 8. The method according to Clause 7 further includes:

[0162] determining a first polarization value by calculating an average of the average light pulse differences for the first set of sampled phases of the fluorescence emitted along the first direction.

[0163] 9. The method according to Clause 8 further includes:

[0164] The second polarization value is determined by calculating the average value of the average light pulse difference of the second set of sampling phases of the fluorescence emitted along the second direction.

[0165] 10. The method according to claim 9, further comprising:

[0166] determining a concentration value by subtracting the second polarization value from the first polarization value and then dividing by the sum of the first polarization value and the second polarization value.

[0167] 11. The method according to claim 1, wherein the antibody is immunoglobulin G (IgG).

[0168] 12. A fluorescence polarization system for measuring antibody concentration in a sample, the system comprising:

[0169] a light source;

[0170] a first polarization filter that polarizes light emitted from the light source along a first polarization direction;

[0171] a container containing a sample mixed with a fluorescence polarization reagent, the container receiving light polarized along the first direction;

[0172] movable second polarization filters and third polarization filters, the second polarization filter restricting fluorescence emitted from the container to pass through in the first direction, and the third polarization filter restricting fluorescence emitted from the container to pass through in a second direction substantially perpendicular to the first direction;

[0173] a detector for measuring fluorescence emitted along the first direction and fluorescence emitted along the second direction; and

[0174] a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to:

[0175] A measurement cycle is performed, measuring a first set of sampling phases of the fluorescence emitted along the first direction and a second set of sampling phases of the fluorescence emitted along the second direction, wherein at least one sampling phase of the fluorescence emitted along the second direction occurs between sampling phases of the fluorescence emitted along the first direction;

[0176] A first value of the first set of sampling phases is calculated;

[0177] A second value of the second set of sampling phases is calculated; and

[0178] The antibody concentration is determined based on a function of the first value and the second value.

[0179] 13. The system according to claim 12, wherein a first sampling phase of the first set of sampling phases of the fluorescence emitted along the first direction occurs before the midpoint of the measurement cycle, and a second sampling phase of the first set of sampling phases of the fluorescence emitted along the first direction occurs after the midpoint of the measurement cycle.

[0180] 14. The system according to claim 13, wherein each sampling phase includes a voltage measurement of fluorescence from a plurality of light pulses emitted from the light source.

[0181] 15. The system according to Clause 14, wherein, when executed by the processing circuitry system, the instructions further cause the processing circuitry system to:

[0182] obtain a plurality of voltage measurements from each optical pulse.

[0183] 16. The system according to Clause 15, wherein the plurality of voltage measurements from each optical pulse includes a first set of voltage measurements when the light source is turned on and a second set of voltage measurements when the light source is turned off.

[0184] 17. The system according to Clause 16, wherein, when executed by the processing circuitry system, the instructions further cause the processing circuitry system to:

[0185] calculate a first average value of the first set of voltage measurements;

[0186] calculate a second average value of the second set of voltage measurements; and

[0187] determine an optical pulse difference for each optical pulse by subtracting the second average value from the first average value.

[0188] 18. According to the system of Clause 17, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0189] determine an average light pulse difference of the sampled phase by calculating an average of the light pulse differences from the sampled phase.

[0190] 19. According to the system of Clause 18, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0191] determine a first polarization value by calculating an average of the average light pulse differences of the first set of sampled phases of the fluorescence emitted along the first direction.

[0192] 20. According to the system of Clause 19, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0193] determine a second polarization value by calculating an average of the average light pulse differences of the second set of sampled phases of the fluorescence emitted along the second direction.

[0194] 21. According to the system of claim 20, wherein the instruction, when executed by the processing circuit system, further causes the processing circuit system to:

[0195] determine a concentration value by subtracting the second polarization value from the first polarization value and then dividing by the sum of the first polarization value and the second polarization value.

[0196] 22. According to the system of claim 12, wherein the antibody is immunoglobulin G (IgG).

[0197] 23. A method for measuring the concentration of an antibody in a cell sample by fluorescence polarization, the method comprising:

[0198] performing a measurement cycle of fluorescence emitted from within the cell sample mixed with a fluorescence polarization reagent, the measurement cycle comprising the following measurement sequence:

[0199] (1) measuring a first sampling phase of fluorescence emitted along a first direction prior to the midpoint of the measurement cycle;

[0200] (2) Measuring a second sampling phase of fluorescence emitted along a second direction prior to the midpoint of the measurement period, the second direction being perpendicular to the first direction;

[0201] (3) Measuring a third sampling phase of fluorescence emitted along the second direction after the midpoint of the measurement period;

[0202] (4) Measuring a fourth sampling phase of fluorescence emitted along the first direction after the midpoint of the measurement period:

[0203] Calculating a first average value of the first sampling phase and the fourth sampling phase;

[0204] Calculating a second average value of the second sampling phase and the third sampling phase; and

[0205] Determining the antibody concentration based on a function of the first average value and the second average value.

[0206] 24. The method according to claim 23, wherein each sampling phase includes a voltage measurement of fluorescence from a plurality of light pulses emitted from a light source.

[0207] 25. The method according to claim 24, further comprising:

[0208] Obtaining a plurality of voltage measurements from each light pulse.

[0209] 26. According to the method of clause 25, wherein the plurality of voltage measurements from each optical pulse includes a first set of voltage measurements when the light source is turned on and a second set of voltage measurements when the light source is turned off.

[0210] 27. According to the method of clause 26, further comprising:

[0211] calculating an average of the first set of voltage measurements;

[0212] calculating an average of the second set of voltage measurements; and

[0213] determining an optical pulse difference for each optical pulse by subtracting the average of the second set of voltage measurements from the average of the first set of voltage measurements.

[0214] 28. According to the method of clause 27, further comprising:

[0215] determining an average voltage value for each sampling phase by calculating an average of the optical pulse differences of the plurality of optical pulses in each sampling phase.

[0216] 29. According to the method of clause 28, further comprising:

[0217] determining a first polarization value of the first set of measurements for the sampling phase by calculating an average of the average voltage values ​​of the first set of measurements for the sampling phase.

[0218] 30. The method according to Clause 29 further includes:

[0219] determining a second polarization value of the second set of measurements of the sampling phase by calculating the average value of the average voltage values ​​of the second set of measurements of the sampling phase.

[0220] 31. The method according to Clause 30 further includes:

[0221] determining a concentration value by subtracting the second voltage value from the first voltage value and then dividing by the sum of the first voltage value and the second voltage value.

[0222] 32. The method according to Clause 23, wherein the antibody is immunoglobulin G (IgG).

[0223] 33. A fluorescence polarization system for measuring the concentration of antibody proteins in a cell sample, the system comprising:

[0224] a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to:

[0225] perform a measurement cycle of fluorescence emitted from the cell sample, the measurement cycle comprising the following measurement sequence:

[0226] (1) measuring a first sampling phase of fluorescence emitted along a first direction before the midpoint of the measurement cycle;

[0227] (2) measuring a second sampling phase of fluorescence emitted along a second direction before the midpoint of the measurement cycle, the second direction being perpendicular to the first direction;

[0228] (3) measuring a third sampling phase of fluorescence emitted along the second direction after the midpoint of the measurement cycle; and

[0229] (4) measuring a fourth sampling phase of fluorescence emitted along the first direction after the midpoint of the measurement cycle;

[0230] calculating a first average value of the first sampling phase and the fourth sampling phase;

[0231] Calculate a second average value of the second sampling phase and the third sampling phase; and

[0232] determine the antibody concentration based on a function of the first average value and the second average value.

[0233] 34. The system according to claim 33, wherein each sampling phase includes voltage measurements of fluorescence from a plurality of light pulses emitted from a light source.

[0234] 35. The system according to claim 34, wherein the instructions, when executed by the processing circuit system, also cause the processing circuit system to:

[0235] obtain a plurality of voltage measurements from each light pulse.

[0236] 36. The system according to claim 35, wherein the plurality of voltage measurements from each light pulse includes a first set of voltage measurements when the light source is turned on and a second set of voltage measurements when the light source is turned off.

[0237] 37. The system according to Clause 36, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0238] calculate the average value of the first set of voltage measurements;

[0239] calculate the average value of the second set of voltage measurements; and

[0240] determine the optical pulse difference of each optical pulse by subtracting the average value of the second set of voltage measurements from the average value of the first set of voltage measurements.

[0241] 38. The system according to Clause 37, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0242] determine the voltage value of each sampling phase by calculating the average value of the optical pulse differences of the plurality of optical pulses in each sampling phase.

[0243] 39. The system according to claim 38, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0244] determine a first polarization value by calculating an average value of the voltage value for each sampling phase of the fluorescence emitted along the first direction.

[0245] 40. The system according to claim 39, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0246] determine a second polarization value by calculating an average value of the voltage value for each sampling phase of the fluorescence emitted along the second direction.

[0247] 41. The system according to claim 40, wherein, when executed by the processing circuit system, the instructions further cause the processing circuit system to:

[0248] determine the antibody concentration by subtracting the second polarization value from the first polarization value and then dividing by the sum of the first polarization value and the second polarization value.

[0249] 42. According to the system described in Clause 33, the antibody is immunoglobulin G (IgG).

[0250] 43. A fluorescence polarization system for measuring the concentration of immunoglobulin G (IgG) in a sample, the system comprising:

[0251] a processing circuit system having a memory for storing instructions, the instructions, when executed by the processing circuit system, causing the processing circuit system to:

[0252] execute a measurement cycle, measuring a first set of sampling phases of fluorescence emitted along a first direction and a second set of sampling phases of fluorescence emitted along a second direction substantially perpendicular to the first direction, at least one sampling phase of the fluorescence emitted along the second direction occurring between sampling phases of the fluorescence emitted along the first direction, each sampling phase in the first set of sampling phases and the second set of sampling phases including a voltage measurement of light from a plurality of light pulses emitted from a light source;

[0253] obtain a plurality of voltage measurements from each light pulse, wherein the plurality of voltage measurements from each light pulse includes a first set of voltage measurements when the light source is turned on and a second set of voltage measurements when the light source is turned off;

[0254] calculate an average of the first set of voltage measurements;

[0255] calculate an average of the second set of voltage measurements;

[0256] The optical pulse difference for each optical pulse is determined by subtracting the average of the second set of voltage measurements from the average of the first set of voltage measurements;

[0257] The voltage value for each sampling phase is determined by calculating the average of the optical pulse differences of the plurality of optical pulses in each sampling phase;

[0258] The first polarization value for the first set of sampling phases is determined by calculating the average of the voltage values ​​of the first set of sampling phases;

[0259] The second polarization value of the second set of sampling phases is determined by calculating the average value of the voltage values ​​in the second set of sampling phases; and

[0260] the concentration value of the IgG is determined by subtracting the second polarization value from the first polarization value and then dividing by the sum of the first polarization value and the second polarization value.

[0261] 44. A system for measuring antibody concentration in a sample, the system comprising:

[0262] a light source;

[0263] an excitation polarizer that polarizes light emitted from the light source along a first direction;

[0264] a container configured to receive light polarized along the first direction;

[0265] a polarizer holder that holds a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from within the container to pass through in the first direction, and the second detection polarizer restricting fluorescence emitted from within the container to pass through in a second direction perpendicular to the first direction;

[0266] a detector for measuring fluorescence emitted along the first direction and fluorescence emitted along the second direction; and

[0267] a processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause the processing circuit system to:

[0268] The polarizer holder is rotated about a rotation axis in alternating directions such that the first detection polarizer and the second detection polarizer are alternately positioned in the optical path of the fluorescence emitted from the container.

[0269] 45. The system according to claim 44, wherein the instruction, when executed by the processing circuit system, further causes the processing circuit system to:

[0270] rotate the polarizer holder to position the first detection polarizer in the optical path of the fluorescence emitted from the container; and

[0271] rotate the polarizer holder to position the second detection polarizer in the optical path of the fluorescence emitted from the container.

[0272] 46. The system according to claim 44, further comprising:

[0273] an optical plate having a groove terminating in the first stop and the second stop; and

[0274] wherein the polarizer holder includes a pin that, when the polarizer holder is rotated about the rotation axis, displaces within the groove of the optical plate.

[0275] 47. According to the system described in Clause 46, the optical plate further includes a first magnet positioned next to the first stop in the groove and a second magnet positioned next to the second stop in the groove; and

[0276] wherein the pin is made of a ferromagnetic material and is positioned at the end of the first stop at the first stop.When the pin is next to the moving part, it is attracted to the first magnet, and when it is positioned next to the second stop at the end of the second rotational stroke, it is attracted to the second magnet.

[0277] 48. The system according to claim 46, wherein the groove is bent along the axis of rotation.

[0278] 49. The system according to claim 44, wherein the polarizer holder includes a first side surface and a second side surface perpendicular to the first side surface, the first side surface including a first recess for receiving the first detection polarizer, and the second side surface including a second recess for receiving the second detection polarizer.

[0279] 50. The system according to claim 49, wherein the polarizer holder further includes a first biasing element attached to the first side surface to secure the first detection polarizer in the first recess, and a second biasing element attached to the second side surface to secure the second detection polarizer in the second recess.

[0280] 51. The system according to Clause 50, wherein when the first detection polarizer and the second detection polarizer are respectively placed in the first recess and the second recess, they protrude beyond the first side surface and the second side surface, and the first bias element and the second bias element are bent around the first detection polarizer and the second detection polarizer to secure the first detection polarizer and the second detection polarizer in place.

[0281] 52. The system according to Clause 51, wherein the first bias element and the second bias element include cutouts to increase the curvature around the first detection polarizer and the second detection polarizer.

[0282] 53. The system according to Clause 49, wherein the polarizer holder further includes a curved surface connecting the first side surface and the second side surface, the curved surface having a hole aligned with a hole in the first recess and the second recess, thereby allowing the fluorescence emitted from the container to pass through the polarizer holder and reach the detector. Specification 20 / 23 pages 24 CN 121399448 A

[0283] 54. The system according to claim 53 further includes:

[0284] an optical cover that at least partially accommodates the detector and the polarizer holder, the optical cover including a curved wall aligned with a curved surface of the polarizer holder to reduce the space between the polarizer holder and the curved wall.

[0285] 55. The system according to claim 44 further includes:

[0286] a work platform;

[0287] a liquid storage module mounted on the work platform, the liquid storage module including:

[0288] a base having an internal volume for accommodating an internal reservoir;

[0289] A main cover, connected to the base, the main cover including a port providing access to the internal reservoir;

[0290] a secondary cover, attached to the main cover, the secondary cover configured to seal the port on the main cover;

[0291] a second motor, attached to the secondary cover; and

[0292] a dispensing system configured to move a probe along three mutually perpendicular axes above the work platform; and

[0293] wherein, when executed by the processing circuitry system, the instructions also cause the processing circuitry system to:

[0294] operate the second motor to open the secondary cover, thereby allowing the probe of the dispensing system to be inserted through the port on the main cover to reach the bottom of the internal reservoir to draw up the liquid contained therein; and

[0295] when the probe is removed from the port, operate the second motor to close the secondary cover to seal the port on the main cover.

[0296] 56. A polarizer holder for a cell analysis system, the polarizer holder comprising:

[0297] a first side surface;

[0298] a second side surface perpendicular to the first side surface;

[0299] a curved surface connecting the first side surface and the second side surface;

[0300] a first detection polarizer coupled to the first side surface, the first detection polarizer restricting emitted fluorescence to pass in a first direction; and

[0301] a second detection polarizer coupled to the second side surface, the second detection polarizer restricting emitted fluorescence to pass in a second direction;

[0302] the polarizer holder being configured to rotate about a rotation axis such that the first detection polarizer and the second detection polarizer are alternately positioned in the optical path of light.

[0303] 57. The polarizer holder according to claim 56, further comprising:

[0304] a first recess in the first side surface, wherein the first detection polarizer is mounted in the first recess; and

[0305] a second recess in the second side surface, wherein the second detection polarizer is mounted in the second recess.

[0306] 58. The polarizer holder according to claim 57, further comprising:

[0307] a first biasing element attached to the first side surface to secure the first detection polarizer in the first recess; and

[0308] a second biasing element attached to the second side surface to secure the second detection polarizer in the second recess.

[0309] 59. The polarizer holder according to claim 58, wherein when the first detection polarizer and the second detection polarizer are respectively placed in the first recess and the second recess, the polarizer protrudes beyond the first side surface and the second recess.The second side surface, and the first biasing element and the second biasing element are bent around the first detection polarizer and the second detection polarizer to secure the first detection polarizer and the second detection polarizer in place.

[0310] 60. A polarizer holder according to claim 59, wherein the first retaining spring and the second retaining spring include cutouts to increase the curvature around the first detection polarizer and the second detection polarizer.

[0311] 61. A method for measuring the concentration of an antibody in a sample by fluorescence polarization, the method comprising:

[0312] positioning a polarizer holder in a first position such that a first detection polarizer is placed in the optical path of fluorescence emitted from a container, the container containing an antibody sample mixed with a fluorescence polarizing reagent;

[0313] detecting the fluorescence emitted from the container after the light passes through the first detection polarizer, the first detection polarizer restricting the fluorescence emitted from the container to pass through in a first direction;

[0314] rotating the polarizer holder about a rotation axis to position the polarizer holder in a second position such that a second detection polarizer is placed in the optical path of the fluorescence emitted from the container, the second detection polarizer restricting the fluorescence emitted from the container to pass through in a second direction; and

[0315] detecting the fluorescence emitted from the container after the light passes through the second detection polarizer.

[0316] 62. The method according to clause 61 further includes:

[0317] rotating the polarizer holder about the rotation axis to return the polarizer holder from the second position to the first position.

[0318] 63. The method according to clause 61, wherein the polarizer holder is rotated 90 degrees between the first position and the second position.

[0319] 64. The method according to clause 61 further includes:

[0320] determining the antibody concentration based on detecting light passing through the first detection polarizer and the second detection polarizer.

[0321] 65. A module for a cell analysis system, the module comprising:

[0322] a base having an internal volume for receiving an internal reservoir;

[0323] a main cover connected to the base, the main cover including a port providing access to the internal reservoir;

[0324] a secondary cover attached to the main cover, the secondary cover configured to seal the port on the main cover;

[0325] a motor attached to the secondary cover; and

[0326] a processing circuit system having a memory for storing instructions, the instructions, when executed by the processing circuit system, causing the processing circuit system to:

[0327] operate the motor to open the secondary cover, thereby allowing a probe to be inserted through the port on the main cover to...Reaching the bottom of the internal reservoir; and

[0328] when the probe is removed, operating the motor to close the secondary cover thereby sealing the port on the main cover.

[0329] 66. The module according to claim 65, further comprising:

[0330] a gasket that seals around the internal reservoir when the main cover is closed.

[0331] 67. The module according to claim 65, further comprising:

[0332] a gasket that seals the port on the main cover when the secondary cover is closed.

[0333] 68. The module according to claim 65, wherein the secondary cover includes a top portion covering the port of the main cover when the secondary cover is closed, and a hinge extending from the top portion, wherein the hinge is connected to the electrical appliance.

[0334] 69. The module according to Clause 65, wherein the secondary cover includes at least one of a magnet and an iron-containing element, the iron-containing element being attracted to a corresponding element on the main cover to hold the secondary cover in a closed state.

[0335] 70. The module according to Clause 65, wherein the secondary cover includes a plunger configured to be at least partially inserted into the port on the main cover. Instruction manual page 23 / 23, 27 CN 121399448 A, Figure 1; Instruction manual figure 1 / 38, page 28, CN 121399448 A, Figure 2; Instruction manual figure 2 / 38, page 29, CN 121399448 A, Figure 3; Instruction manual figure 3 / 38, page 30, CN 121399448 A, Figure 4; Instruction manual figure 4 / 38, page 31, CN 121399448 A, Figure 5; Instruction manual figure 5 / 38, page 32, CN 121399448 A, Figure 6; Instruction manual figure 6 / 38, page 33, CN 121399448 A, Figure 7; Figure 8; Instruction manual figure 7 / 38, page 34, CN 121399448 A, Figure 9; Instruction manual figure 8 / 38, page 35, CN 121399448 A, Figure 10; Instruction manual figure 9 / 38, page 36, CN 121399448 A, Figure 11. Instruction manual illustrations, page 10 / 38, 37 CN 121399448 A, Figure 12; Instruction manual illustrations, page 11 / 38, 38 CN 121399448 A, Figure 13; Instruction manual illustrations, page 12 / 38, 39 CN 121399448 A, Figure 14; Instruction manual illustrations, page 13 / 38, 40 CN 121399448 A, Figure 15; Description.Figure 16 of the instruction manual, page 41 of page 38 (CN 121399448 A); Figure 17 of the instruction manual, page 43 of page 16 of page 38 (CN 121399448 A); Figure 18 of the instruction manual, page 44 of page 17 of page 38 (CN 121399448 A); Figure 19 of the instruction manual, page 45 of page 18 of page 38 (CN 121399448 A); Figure 20 of the instruction manual, page 46 of page 19 of page 38 (CN 121399448 A); Figure 21 of the instruction manual, page 47 of page 20 of page 38 (CN 121399448 A); Figure 22 of the instruction manual, page 48 of page 21 of page 38 (CN 121399448 A); Figure 23 of the instruction manual, page 49 of page 22 of page 38 (CN 121399448 A); Figure 24 of the instruction manual, page 23 of page 38 (CN 121399448 A); Figure 50 of the instruction manual, page 38 (CN 121399448 A). Figure 25 of the instruction manual, attached to page 24 / 38, CN 121399448 A; Figure 26 of the instruction manual, attached to page 25 / 38, CN 121399448 A; Figure 27 of the instruction manual, attached to page 26 / 38, CN 121399448 A; Figure 28 of the instruction manual, attached to page 27 / 38, CN 121399448 A; Figure 29 of the instruction manual, attached to page 28 / 38, CN 121399448 A; Figure 30 of the instruction manual, attached to page 29 / 38, CN 121399448 A; Figure 31 of the instruction manual, attached to page 30 / 38, CN 121399448 A; Figure 32 of the instruction manual, attached to page 31 / 38, CN 121399448 A; Figure 33 of the instruction manual, attached to page 32 / 38, CN 121399448 A; Figure 59 of the instruction manual, CN 121399448 A. Figure 34 Appendix to the Instruction Manual, Page 33 / 38, 60 CN 121399448 A Figure 35 Figure 36 Appendix to the Instruction Manual, Page 34 / 38, 61 CN 121399448 A Figure 37 Figure 38 Appendix to the Instruction Manual, Page 35 / 38, 62 CN 121399448 A Figure 39 Appendix to the Instruction Manual, Page 36 / 38, 63 CN 121399448 A Figure 40 Appendix to the Instruction Manual, Page 37 / 38, 64 CN 121399448 A Figure 41 Appendix to the Instruction Manual, Page 38 / 38, 65 CN 121399448 A

Claims

1. A system for measuring an antibody concentration in a sample, the system comprising: a light source; an excitation polarizer that polarizes light emitted from the light source in a first direction; a vessel configured to receive light polarized in the first direction; a polarizer holder that holds a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescent light emitted from within the vessel to pass in the first direction, and the second detection polarizer restricting fluorescent light emitted from within the vessel to pass in a second direction, the second direction being perpendicular to the first direction; a detector for measuring fluorescent light emitted in the first direction and fluorescent light emitted in the second direction; and processing circuitry having a memory for storing instructions that, when executed by the processing circuitry, cause the processing circuitry to: rotate the polarizer holder in alternating directions about an axis of rotation such that the first detection polarizer and the second detection polarizer are alternately placed in an optical path of the fluorescent light emitted from within the vessel. the instructions, when executed by the processing circuitry, further cause the processing circuitry to:

2. The system of claim 1, wherein, rotate the polarizer holder to position the first detection polarizer in the optical path of the fluorescent light emitted from within the vessel; and rotate the polarizer holder to position the second detection polarizer in the optical path of the fluorescent light emitted from within the vessel.

3. The system of claim 1, further comprising: an optical plate having a groove that terminates in a first stop and a second stop; and wherein the polarizer holder includes a pin that is displaced within the groove of the optical plate as the polarizer holder is rotated about the axis of rotation. the optical plate further includes a first magnet positioned alongside the first stop of the groove and a second magnet positioned alongside the second stop of the groove; and wherein the pin includes a ferromagnetic material that is attracted to the first magnet when positioned alongside the first stop at the end of a first rotation stroke and is attracted to the second magnet when positioned alongside the second stop at the end of a second rotation stroke.

4. The system of claim 3, wherein, the groove is curved along the axis of rotation. the polarizer holder includes a first side surface and a second side surface that is perpendicular to the first side surface, the first side surface including a first recess for receiving the first detection polarizer and the second side surface including a second recess for receiving the second detection polarizer.

5. The system of claim 3, wherein, the polarizer holder further includes a first biasing element attached to the first side surface to secure the first detection polarizer in the first recess, and a second biasing element attached to the second side surface to secure the second detection polarizer in the second recess.

6. The system of claim 1, wherein, ​ 7. The system of claim 6, wherein, ​ 8. The system of claim 7, wherein, When the first and second detection polarizers are placed within the first and second recesses, respectively, the first and second detection polarizers protrude beyond the first and second side surfaces, and the first and second biasing elements are bent around the first and second detection polarizers to secure the first and second detection polarizers in place.

9. The system of claim 8, wherein, The first and second biasing elements include cutouts to increase the degree of bending around the first and second detection polarizers.

10. The system of claim 6, wherein, The polarizer holder further includes a curved surface connecting the first and second side surfaces, the curved surface having an aperture aligned with the aperture in the first and second recesses, thereby allowing the emitted fluorescence to pass through the polarizer holder from within the container and to the detector.

11. The system of claim 10, further comprising: an optical cover at least partially housing the detector and the polarizer holder, the optical cover including a curved wall aligned with the curved surface of the polarizer holder to reduce the space between the polarizer holder and the curved wall.

12. The system of claim 1, further comprising: a work platform; a liquid storage module mounted on the work platform, the liquid storage module comprising: a base having an internal volume for housing an internal reservoir; a main cover connected to the base, the main cover including a port providing access to the internal reservoir; a secondary cover attached to the main cover, the secondary cover configured to seal the port on the main cover; a second motor attached to the secondary cover; and a dispensing system configured to move a probe above the work platform along three mutually perpendicular axes; and wherein the instructions, when executed by the processing circuitry, further cause the processing circuitry to: operate the second motor to open the secondary cover, thereby allowing a probe of the dispensing system to be inserted through the port on the main cover to reach a bottom of the internal reservoir to aspirate a liquid housed in the internal reservoir; and operate the second motor to close the secondary cover to seal the port on the main cover when the probe is removed from the port.

13. A polarizer holder for a cell analysis system, the polarizer holder comprising: a first side surface; a second side surface perpendicular to the first side surface; a curved surface connecting the first and second side surfaces; a first detection polarizer coupled on the first side surface, the first detection polarizer restricting emitted fluorescence to pass in a first direction; and a second detection polarizer coupled on the second side surface, the second detection polarizer restricting emitted fluorescence to pass in a second direction; the polarizer holder configured to rotate around an axis of rotation, alternating the first and second detection polarizers to be placed in the optical path of the light.

14. The polarimeter holder of claim 13, further comprising: a first recess in the first side surface, wherein the first detection polarimeter is mounted in the first recess; and a second recess in the second side surface, wherein the second detection polarimeter is mounted in the second recess.

15. The polarimeter holder of claim 14, further comprising: a first biasing element attached to the first side surface to secure the first detection polarimeter in the first recess; and a second biasing element attached to the second side surface to secure the second detection polarimeter in the second recess.

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