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

Cross-references to related applications

[0001] This application was filed on July 19, 2024 as a PCT international application 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 STORAGEMODULE FOR CELL ANALYSIS SYSTEM,” the disclosure of which is incorporated herein by reference in its entirety. Background Technology

[0002] Antibodies are large, Y-shaped proteins used by the immune system to recognize and neutralize foreign substances, such as pathogenic bacteria and viruses. Antibodies recognize unique molecules (called antigens) of pathogens. Each tip of a Y-shaped antibody contains a complementary site that matches an epitope on the antigen, allowing the antibody to bind to the antigen. Using this binding mechanism, antibodies can tag microbes or infected cells for attack by other parts of the immune system, or they can directly neutralize microbes 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 using fluorescence polarization. Summary of the Invention

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

[0004] One aspect relates to a method for measuring the concentration of an antibody in a sample using fluorescence polarization, 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 the fluorescence emitted along the second direction occurring between the sampling phases of the fluorescence emitted along the first direction; calculating a first value of the first set of sampling phases; calculating a second value of the second set of sampling phases; and determining an 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 disclosed, the system comprising: a light source; a first polarization filter that polarizes light emitted from the light source along a first direction; a container holding a sample mixed with a fluorescent polarizing reagent, the container receiving light polarized along the first direction; movable second 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; and 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 antibody concentration in a cell sample by fluorescence polarization is provided, the method comprising: performing a measurement cycle of fluorescence emitted from within the 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 along a first direction before the midpoint of the measurement cycle; (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; (3) measuring a third sampling phase of fluorescence emitted along the second direction after the midpoint of the measurement cycle; and (4) measuring a fourth sampling phase of fluorescence emitted along the first direction after the midpoint of the measurement cycle; 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.

[0007] On the other hand, a fluorescence polarization system for measuring the concentration of antibody proteins in a cell sample is involved. 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 comprising the following measurement sequence: (1) measuring a first sampling phase of fluorescence emitted along a first direction before the midpoint of the measurement cycle; (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; (3) measuring a third sampling phase of fluorescence emitted along the second direction after the midpoint of the measurement cycle; and (4) measuring a fourth sampling phase of fluorescence emitted along the first direction after the midpoint of the measurement cycle; 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 and second averages.

[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; a detector for measuring fluorescence emitted along the first direction and fluorescence emitted along the 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 within the container.

[0010] On the other hand, a polarizer holder for a cell analysis system is disclosed, 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 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 detection polarizer and the second detection polarizer 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 disclosed, the module comprising: a base having an internal volume for accommodating 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 and 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 involve individual features and combinations of features. It should be understood that both the foregoing general description and the following detailed description are exemplary and illustrative only, and do not limit the broad inventive concept on which the embodiments disclosed herein are based. Attached Figure Description

[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] Figure 1 This is an isometric view of an example of a cell analysis system for analyzing cell health in multiple cell samples.

[0016] Figure 2 yes Figure 1 Another isometric view of the cell analysis system with the top cover removed from the housing.

[0017] Figure 3 yes Figure 1 A top view of the working platform supported inside the housing of the cell analysis system.

[0018] Figure 4 schematically shown Figure 3 An example of a titer module supported on a working platform.

[0019] Figure 5 The diagram is shown graphically. Figure 4 An example of a graph showing the error in calculating antibody concentration caused by detected light intensity drift in the titer module.

[0020] Figure 6 The diagram is shown graphically by [the following text is incomplete and likely refers to a separate topic:] Figure 5 The curve shown is a graph of the simulated fluorescence polarization measurement error generated by the measurement sequence.

[0021] Figure 7 It schematically shows that it can be made by Figure 4An example of a method for measuring antibody concentration in a cell sample, executed by the titer module.

[0022] Figure 8 It schematically shows that in Figure 7 This is an example of a method that performs a measurement cycle during the operation of a method.

[0023] Figure 9 It schematically shows the following based on Figure 8 An example of a measurement cycle performed using the method described above.

[0024] Figure 10 The diagram is shown graphically. Figure 7 An example of a graph illustrating how the method reduces the error in calculating antibody concentration in cell samples.

[0025] Figure 11 The diagram is shown graphically by [the following text is incomplete and likely refers to a separate topic:] Figures 8 to 10 The graph shows the reduction in fluorescence polarization measurement error caused by the measurement cycle.

[0026] Figure 12 It schematically shows that in Figures 8 to 10 The method of measuring the sampling phase during the measurement cycle.

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

[0028] Figure 14 It is possible Figure 1 An isometric view of an example of a titer module used in a cell analysis system.

[0029] Figure 15 yes Figure 14 An isometric view of the titer module, with a portion of the housing removed to expose the internal components of the titer module.

[0030] Figure 16 It is along Figure 14 An equidistant cross-sectional view taken from the horizontal axis of the titer module.

[0031] Figure 17 yes Figure 14 An isometric view of the titration module, with the inner cover removed to expose the optical components of the titration module.

[0032] Figure 18 yes Figure 14 Right side view of the optical components of the titration module

[0033] Figure 19 yes Figure 14Left-side view of the optical components of the titration module.

[0034] Figure 20 It is along Figure 14 An equidistant cross-sectional view taken from the vertical axis of the titer module.

[0035] Figure 21 yes Figure 14 The right-side view of the cross-section of the titer module.

[0036] Figure 22 yes Figure 14 The left-side cross-section view of the titer module.

[0037] Figure 23 yes Figure 14 Isometric view of the polarizer holder, optical plate, and detector of the titration module.

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

[0039] Figure 25 yes Figure 14 An exploded view of the polarizer holder of the titer module.

[0040] Figure 26 yes Figure 14 An exploded view of the polarizer holder of the titer module.

[0041] Figure 27 yes Figure 14 A top view of the polarizer holder of the titration module.

[0042] Figure 28 yes Figure 14 An isometric view of the optical plate and motor of the titration module, with the polarizer holder removed from it.

[0043] Figure 29 yes Figure 26 A top view of the optical plate.

[0044] Figure 30 yes Figure 1 An isometric view of the liquid storage module of the cell analysis system.

[0045] Figure 31 yes Figure 30 Another isometric view of the liquid storage module shows the lid of the liquid storage module in the open position.

[0046] Figure 32 yes Figure 30 Another isometric view of the liquid storage module shows the internal storage unit removed from the interior of the liquid storage module.

[0047] Figure 33 yes Figure 30 Another isometric view of the liquid storage module shows the automatic cover of the liquid storage module in the open position.

[0048] Figure 34 yes Figure 30 Another isometric view of the liquid storage module shows the lid of the liquid storage module in the open position.

[0049] Figure 35 yes Figure 30 An isometric view of the sub-cap of the liquid storage module.

[0050] Figure 36 yes Figure 35 Front view of the secondary cover.

[0051] Figure 37 yes Figure 35 Side view of the secondary cover.

[0052] Figure 38 yes Figure 35 A bottom view of the sub-cover.

[0053] Figure 39 yes Figure 1 An isometric view inside the cell analysis system, showing the dispensing system positioning the probe. Figure 30 Near the liquid storage module.

[0054] Figure 40 yes Figure 1 An isometric view inside the cell analysis system, showing the dispensing system inserting probes. Figure 30 The liquid storage module contains internal storage.

[0055] Figure 41 It shows Figure 14 An example of an alternative implementation of the polarizer holder for the titer module. Detailed Implementation

[0056] Figure 1 This is an isometric view of an example of a cell analysis system 100 for analyzing cell health in 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 to automated bioreactors and other systems and devices. This connectivity may include electrical connections for sharing data and physical connections for liquid handling purposes. The cell analysis system 100 automates sample preparation and minimizes the sample volume required to measure cell characteristics. The 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] like Figure 1 As shown, the cell analysis system 100 includes a housing 102 that supports a working platform 300. The housing 102 includes a top cover 104, which can support the reference... Figure 2 and Figure 3 The allocation system 106 is described in more detail.

[0059] Figure 2 This is another isometric view of the cell analysis system 100, in which the top cover 104 is removed from the housing 102, thereby exposing the dispensing system 106. Figure 3 This is a top view of the working platform 300 supported inside the housing 102. Now refer to... Figure 2 and Figure 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 for liquid aspiration and dispensing.

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

[0062] like Figure 2As shown, housing 102 is sized to have a width W, a depth D, and a height H. As an illustrative example, the width W is approximately 24 inches to approximately 36 inches, the depth D is approximately 24 inches to approximately 36 inches, and the height H is approximately 24 inches to approximately 36 inches. In some examples, housing 102 is sized to have a cubic shape.

[0063] like Figure 3 As shown, the work platform 300 supports a sample transfer module 302, one or more mixing plates 304, 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 refer to... Figure 4 To describe in more detail.

[0064] Figure 4 An example of a titer module 400 supported on a work platform 300 is schematically shown. The titer module 400 includes optical components for measuring the concentration of antibodies present in a cell sample. For example, the titer module 400 measures the concentration of immunoglobulin G (IgG) present in a cell sample.

[0065] More specifically, the titer module 400 calculates fluorescence polarization measurements to measure the concentration of IgG protein in a cell sample. The fluorescence polarization method involves mixing the cell sample with a fluorescence polarization 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 techniques described herein can be used to measure the concentrations of other types of proteins and antibodies.

[0066] like Figure 4 As shown, the titration module 400 includes a light source 402 that emits light 404 into a container 412 containing a cell solution mixed with a 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 container 412, eliminating the need for the user of the system to manually mix the solution in container 412. In some examples, container 412 is a cuvette, test tube, etc.

[0067] Light source 402 emits light 404 without polarization, such that light 404 is unpolarized light. In some examples, 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 the light 404 emitted from light source 402, respectively.

[0068] Light 404 passes through an excitation polarizer 410, which polarizes the light 404 along a first direction 411. In some examples, the first direction 411 is a linear direction. Figure 4 In the example shown, 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 light 404 is polarized along the first direction 411, it is absorbed by the cell solution in container 412, which is mixed with the fluorescent polarizing reagent. 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 the 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 polarized fluorescence 416 to pass through in the second direction 413. Thereafter, the polarized fluorescence 416 passes through a spectral filter 424 before being detected by the detector 426. In some examples, the detector 426 is a photomultiplier tube (PMT). The detector 426 converts the 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] Where I PAR The voltage of the parallel fluorescence detected from the solution in container 412, I PERFP is the voltage of the vertical fluorescence detected from the solution in container 412, and FP is the fluorescence polarization in millipoles (mP). As an example, I is detected by detector 426. PAR and I PER The voltage range is from about 0.02 volts to about 5.0 volts.

[0075] The measured fluorescence polarization (FP) is correlated with the antibody concentration in the cell sample. For example, a higher detected FP is associated with a higher antibody concentration, while a lower detected FP is associated with a lower antibody concentration. This is because a higher antibody concentration increases binding to the fluorescent polarizing agent, resulting in a higher voltage for parallel fluorescence and a lower voltage for perpendicular fluorescence, which leads to a higher detected FP.

[0076] The technical challenge of Equation 1 lies in measuring I PAR Any error in time (in I) PER The absence of certain parameters (such as those not present in the original text) can lead to errors in fluorescence polarization (FP) calculations. For example, FP calculations depend on the measured ratio I. PER / I PAR Any proportional impact on I PAR and I PER Changes will be canceled out. However, in measurements such as parallel fluorescence (I0), PAR ) and vertical fluorescence (I PER Signal intensity changes during the process and their impact on vertical fluorescence (I) PER ) and parallel fluorescence (I PAR The offset errors of the two will not be canceled out and will cause significant errors in FP calculation, and thus cause 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 the light source 402, electrical response, dye bleaching, and non-uniform fluid and diffusion in container 412. Other 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 An example of a graph 500 is shown graphically, illustrating the error in FP calculation caused by light intensity drift detected in titration module 400 and the incorrect calculation of antibody concentration in the cell sample. Graph 500 includes light intensity (Y-axis) measured by detector 426 over time (X-axis). In this example, the light intensity was measured without fluorescent polarizing reagent 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, IPER Fluorescence and I PAR The ratio between 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 I PER Fluorescence and I PAR Error in the ratio between fluorescence. In this example, a measurement cycle is performed, in which I is measured in the first phase (i.e., using the second detection polarizer 422). PAR Fluorescence, and subsequently I measured in the second phase (i.e., using the first detection polarizer 420). PAR Fluorescence. I PER fluorescence and I PAR Each fluorescence phase includes two measurements. For example, I PER Fluorescence includes a first measurement of 3.60 V and a second measurement of 3.62 V, and I PAR Fluorescence was measured at a third value of 3.64 V and a fourth value of 3.66 V. I PER Fluorescence was calculated as the average of the first and second measurements (i.e., 3.61 V), and I... PAR Fluorescence was calculated as the average of the third and fourth measurements (i.e., 3.65 V). This resulted in I... PER Fluorescence and I PAR The ratio between the fluorescence particles is 3.61:3.65, which is an error of about 1% caused by the drift of the light source 402.

[0081] Figure 6 The diagram is shown graphically by [the following text is incomplete and likely refers to a separate topic:] Figure 5 The curve of the simulated fluorescence polarization measurement error generated by the measurement sequence is shown in Figure 600. Figure 6 As shown, for lower mP values, such as less than 200 units, the measurement error is significantly larger. Therefore, lower concentrations of antibody in the sample contained in the container are particularly sensitive to changes in signal intensity and noise in the titer module 400.

[0082] In some cases, by implementing sophisticated circuit design and temperature-controlled light sources and detectors within the titration module, Ig from signal strength variations and noise is reduced. PER Fluorescence and I PAR Measurement error in the ratio between fluorescence. However, such a solution is expensive, unreliable, may increase the size of the titration module, and slow down the warm-up time of the device.

[0083] Figure 7 An example of a method 700 for measuring antibody concentration in a cell sample is illustrated schematically. Method 700 uses optical components of the titer module 400 (such as...). Figure 4 (As shown) 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 the dispensing system 106 of the 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 the titer module 400. In effect, Method 700 is a software solution that improves the functionality of the titer module 400 by reducing these types of errors to increase the accuracy of antibody concentration calculations.

[0085] Method 700 includes an operation 702 of 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 the titration 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 with each other.

[0086] Figure 8 An example of method 800 performing a measurement cycle in operation 702 of method 700 is illustrated schematically. Figure 9 An example of a measurement cycle 900 performed according to method 800 is illustrated schematically.

[0087] Now refer to Figure 8 and Figure 9 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 period 900. Next, method 804 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 period 900.

[0088] like Figure 8 and Figure 9 As further shown, 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] The measurement period 900 includes at least one sampling phase along a first direction 411 and at least one sampling phase along a second direction 413 before the midpoint. The measurement period also 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 the measurement period.

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

[0091] like Figure 9 As further shown, each of the sampling phases A to D includes a voltage measurement of the fluorescence from a plurality of light pulses 908 emitted by 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 includes a first set of voltage measurements 910a when the light source 402 is turned on and a second set of voltage measurements 910b when the light source 402 is turned 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 may each include performing Figure 12 The additional operations shown are illustrated.

[0092] Figure 12 It schematically shows that in Figures 8 to 10 The method 1200 for measuring the sampling phase during the measurement cycle. Now refer to Figure 9 and Figure 12 Method 1200 includes an operation 1202 of calculating the average value of a first set of voltage measurements 910a when the light source 402 is turned on. Next, method 1200 includes an operation 1204 of calculating the average value of a second set of voltage measurements 910b when the light source 402 is turned 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] Return to reference Figure 7 Method 700 includes an operation 704 of calculating a first average value of the first sampling phase A and the fourth sampling phase D. For example... Figure 9 As shown, the first sampling phase A includes fluorescence emitted along the 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 the second sampling phase B and the third sampling phase C. As described above, the second sampling phase B includes fluorescence emitted from the container along the second direction 413 before the midpoint 902 of the measurement period 900. The third sampling phase C includes fluorescence emitted from the container along the second direction 413 after the midpoint 902 of the measurement period 900.

[0095] Method 700 includes an operation 708 of determining 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 (IL). PAR The second average value is the voltage of vertical fluorescence (I). PER And use Formula 1 to calculate the antibody concentration.

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

[0097] like Figure 10 As shown, during the measurement cycle, the first sampling phase A for fluorescence emitted along the first direction before the midpoint M is (3.55 V), the second sampling phase B for fluorescence emitted along the second direction before the midpoint M is (3.59 V), the third sampling phase C for fluorescence emitted along the second direction after the midpoint M is (3.61 V), and the fourth sampling phase D for fluorescence emitted along the first direction after the midpoint M is (3.65 V). The first average of the first sampling phase A and the fourth sampling phase D is calculated to be 3.6 V. The second average of the second sampling phase B and the third sampling phase C is calculated to be 3.6 V. This results in I PER Fluorescence and I PAR The ratio of fluorescence is 3.6:3.6, which eliminates errors caused by the drift of light source 402 (see [link]). Figure 5(Compare with curve 500 shown). 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 The diagram is shown graphically by [the following text is incomplete and likely refers to a separate topic:] Figures 8 to 10 The curve 1100 shows the reduction in the simulated fluorescence polarization measurement error caused by the measurement period. Now refer to... Figure 6 and Figure 11 Graph 1100 shows that for lower mP values ​​(e.g., less than 200 units), the measurement error is significantly smaller than that shown in Graph 600 when performing conventional measurement sequences. Graph 1100 also shows that for lower antibody concentrations, the sensitivity to signal intensity variations and noise in the titer module 400 is significantly reduced.

[0099] Figure 13 An example of a controller 1300 of a cell analysis system 100 is schematically shown, which can be used to implement the aspects described herein, including the features of the titer module 400. As... Figure 13 As shown, controller 1300 includes one or more processing devices 1302, memory storage device 1304, and system bus 1306 coupling 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] like Figure 13 As shown, memory storage device 1304 may include random access memory (“RAM”) 1308 and read-only memory (“ROM”) 1310. Basic input and output logic having basic routines that facilitate the transfer of information between elements within controller 1300 (e.g., during startup) may be stored in 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 computer-readable data storage medium contained herein refers to mass storage device 1312, those skilled in the art will understand that a 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. A computer-readable storage medium can consist entirely of non-transitory media. Mass storage device 1312 is an example of a computer-readable storage device.

[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. Examples 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 is accessible by a device.

[0104] Controller 1300 can operate in a networked environment using logical connections to other devices via network 1320. Controller 1300 connects to network 1320 via network interface unit 1318 connected to system bus 1306. 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). Network interface unit 1318 can connect controller 1300 to additional networks, systems, and devices. Controller 1300 also includes an input / output unit 1322 for receiving and processing inputs and outputs from peripheral devices.

[0105] 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. Mass storage device 1312 and / or RAM 1308 may also store software instructions and data 1216, which, when executed by processing device 1302, provide the functionality of the cell analysis system 100 discussed herein.

[0106] Figure 14 This is an isometric view of an example of a titer module 400 that can be used in the cell analysis system 100 (see also, for example, see also). Figure 3 and Figure 4 ). Figure 15 This is an isometric view of the titer module 400, with a portion of the housing 401 removed to expose the internal components of the titer module. Figure 16 This is an equidistant cross-sectional view taken along the horizontal axis of the titer module 400. Now refer to... Figures 14 to 16The internal components of the titration module 400 include an excitation component 403 and a detection component 405.

[0107] The excitation assembly 403 includes a first optical cover 407 that at least partially houses the light source 402 and the excitation polarizer 410. The detection assembly 405 includes a second optical cover 409 that at least partially houses the detector 426, the polarizer holder 418, the first detection polarizer 420, and the 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 typically 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] Figure 17 This is an isometric view of the titration module 400, in which the first optical cover 407 and the second optical cover 409 have been removed, exposing the optical components of the titration module. Figure 18 This is a right-side view of the optical components of the titration module 400. Figure 19 This is a left-side view of the optical components of the titration module 400. Figures 17 to 19 The optical components of the titration module 400 shown are substantially similar to Figure 4 The optical components are schematically shown. For example, the titration module 400 includes a light source 402 that emits light into a container 412 containing a cell culture of IgG protein mixed with a fluorescent polarizing reagent. In some examples, the container 412 is a cuvette, test tube, etc.

[0110] Light source 402 emits light without polarization, making the light unpolarized. In some examples, 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 light source 402, respectively.

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

[0112] Polarized fluorescence, after being emitted from container 412, 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 polarizer holder 418. As will be described in more detail, polarizer holder 418 is powered by 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 the polarized fluorescence into a voltage value to be input into Equation 1 to determine the antibody concentration produced by the cell culture.

[0115] Figure 20 It is an equidistant cross-sectional view taken along the vertical axis of the titration module 400. Figure 21 This is a right-side cross-sectional view of the titer module 400. Figure 22 This is a left-side cross-sectional view of the titer module 400. Now refer to... Figures 20 to 22 The polarizer holder 418 is connected to the drive shaft 434 of the motor 428. In some examples, the motor 428 is a stepper motor (also called a stepper motor or stepper motor) or a similar type of electric motor. In other examples, the motor 428 is a solenoid. The motor 428 is mounted to the underside of the optical plate 419 that supports the optical components of the titration module 400.

[0116] Controller 1300 controls motor 428 to rotate polarizer holder 418 about rotation axis RR in clockwise direction D1 and counterclockwise direction D2 (see...) Figure 27 This allows the first detection polarizer 420 and the second detection polarizer 422 to be alternately positioned in the optical path of the polarized fluorescence emitted from the solution in container 412 before the light reaches detector 426. In some examples, controller 1300 controls motor 428 to rotate polarizer holder 418 about 90 degrees clockwise D1 and about 90 degrees counterclockwise D2 about rotation axis RR.

[0117] In alternative implementation methods Figure 41An example of a polarizer holder 418a is shown, which includes a swivel mount 462 and an electromagnetic actuator 464, which can be used to position a detection polarizer 466 in the path of fluorescence emitted from a container 412. In some examples, the electromagnetic actuator 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 rotatable mount 462, and the magnet 468 drives the rotatable 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 rotatable 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] Figure 23 and Figure 24 This is an isometric view of the polarizer holder 418, optical plate 419, and detector 426 of the titration module 400. Figure 25 and Figure 26 This is an exploded view of the polarizer holder 418. Figure 27 This is a top view of the polarizer holder 418. Now refer to... Figures 23 to 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 in which the first detection polarizer 420 is placed. Similarly, the second side surface 432 includes a recess 435 in which the second detection polarizer 422 is placed. Figure 27As shown, 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] like Figures 25 to 27 As shown, the biasing element 436 is used to hold the first detection polarizer 420 and the second detection polarizer 422 within recesses 435 in the first side surface 430 and the second side surface 432, respectively. A fastener 438, such as a screw, can be screwed through a hole 440 in the biasing element 436 into a hole 441 in the first side surface 430 and the second side surface 432 to attach the biasing 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 recesses 435. In some examples, the biasing element 436 is a retaining spring.

[0121] The biasing 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 hold the first detection polarizer 420 and the second detection polarizer 422 in place. In some examples, the biasing elements 436 include cutouts 439 to increase their flexibility. In some examples, the cutouts 439 are positioned towards the top and bottom of each biasing element 436 to increase the flexibility of the biasing 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. Each recess 435 includes an aperture 437 (see...). Figure 25 The hole 437 corresponds to the corresponding hole 444 on the curved surface 431 (see...). Figure 26 Alignment is achieved, thereby allowing polarized fluorescence to pass through the polarizer holder 418 and reach the detector 426.

[0123] like Figure 16 As shown, 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] Figure 28This is an isometric view of the optical plate 419 and motor 428 of the titration module 400, with the polarizer holder removed from it. Figure 29 This is a top view of optical plate 419. Now refer to... Figure 28 and Figure 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. When the polarizer holder 418 rotates about the rotation axis RR in clockwise direction D1 and counterclockwise direction D2 (see...), Figures 20 to 22 as well as Figure 27 The positioning pin 446 slides within the groove 448 of the optical plate 419. For example... Figure 28 and Figure 29 As shown, groove 448 bends along the rotation axis RR.

[0125] The locating pin 446 contacts the first stop 450 and the second stop 451 in the groove 448 at the end of each rotational stroke in the clockwise direction D1 and the counterclockwise direction D2. For example, when the polarizer holder 418 rotates 90 degrees clockwise D1 to position the first detection polarizer 420 in the optical path, the locating 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 rotates 90 degrees counterclockwise D2 to position the second detection polarizer 422 in the optical path, the locating pin 446 contacts the second stop 451 at the relatively 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 [link to original text]). Figure 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] like Figure 28 and Figure 29 As further shown, the optical plate 419 includes a magnet 452 positioned next to each of the first stop 450 and the second stop 451. A positioning pin 446 is made of a ferromagnetic material such as steel, such that the magnet 452 attracts the positioning pin 446 when positioned near the first stop 450 at the end of a rotational stroke in the clockwise direction D1, and when positioned near the second stop 451 at the end of a rotational stroke in the counterclockwise direction D2.

[0128] The controller 1300 is programmed to shut off the motor 428 at the end of the rotational stroke in both 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] Figure 30 This is an isometric view of the liquid storage module 314 of the cell analysis system 100 (see also...) Figure 3 ).exist Figure 30 In the diagram, the liquid storage module 314 is shown in the closed position. The liquid storage module 314 can be used in conjunction 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 those 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% (by volume) for an onboard storage lifetime of about 31 days within the cell analysis system 100.

[0131] Figure 31 and Figure 32 This is an isometric view showing the liquid storage module 314 in the open position. Now refer to Figures 30 to 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 cover 3008 can be manually pivoted around hinge 3012 to be in the closed position. Figure 30 ) and opening position ( Figure 31 and Figure 32 The liquid storage module 314 includes a gasket 3014 surrounding an 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 gasket 3014 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 to the closed position about the hinge 3012, the attraction of the one or more first magnets 3018 to the one or more second magnets 3020 can keep the cover closed.

[0135] Figure 33 This is another isometric view of the liquid storage module 314, showing the main cover 3008 in the closed position. Now refer to... Figure 30 and Figure 33 The liquid storage module 314 includes a secondary cover 3010 attached to the main cover 3008. The secondary cover 3010 is controlled by the controller 1300 to open and close as needed by the probes, so that the probes 108 of the dispensing system 106 (see [link to dispensing system]) can be accessed. Figure 2 The controller 1300 approaches the internal reservoir 3006. For example, the controller 1300 opens the subcap 3010 during pipetting and closes the subcap 3010 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] like Figure 33 As shown, when the secondary cap 3010 is opened, port 3022 on the main cap 3008 is exposed. The probe 108 of the dispensing system 106 can be inserted through port 3022 to access the internal reservoir 3006 without opening the main cap 3008. In some examples, port 3022 is accessed by a 55 μL or 230 μL tapered pipette tip. Such pipettes are generally not compatible with solid diaphragm caps.

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

[0138] Figure 34 This is another isometric view of the liquid storage module 314, showing the main cover 3008 in the open position. Now refer to... Figure 34 A magnet 3026 is mounted to the main cover 3008, which attracts an iron-containing element 3028 on the secondary cover 3010 to close the secondary cover 3010. For example, the hinge 3030 of the secondary cover 3010 may include the iron-containing element 3028 attracted to the magnet 3026 to close the secondary cover 3010. In an alternative example, the magnet 3026 is included on the secondary cover 3010, and the iron-containing element 3028 is included on the main cover 3008. The magnet 3026 and the iron-containing element 3028 keep the secondary 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 may include a stepper motor or a similar type of electric motor. Motor 3032 is controlled by controller 1300 to move sub-cover 3010 between an open state and a closed state. Motor 3032 may utilize an integrated encoder to track the open and closed states of sub-cover 3010.

[0140] Figure 35 This is an isometric view of the 3010 sub-cover. Figure 36 This is the front view of the sub-cover 3010. Figure 37 This is a side view of the sub-cover 3010. Figure 38 This is a bottom view of the 3010 sub-cover. Now refer to... Figures 35 to 38 The secondary cover 3010 includes a top portion 3040 and a hinge 3042 extending from the top portion. When the secondary cover 3010 is in the closed state, the top portion 3040 covers the port 3022. The hinge 3042 is connected to a motor 3032. For example, the hinge 3042 includes a first hole 3044 which can be used to connect the secondary 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) to a magnet 3026 on the main cover 3008 to keep the secondary cover 3010 closed.

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

[0142] Figure 39 This is an isometric view inside the cell analysis system 100, showing the dispensing system 106 positioning the probe 108 behind the liquid storage module 324. Figure 40 This 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 cap 3010 in the open position. The liquid storage module 324 is designed with 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, allowing the probe 108 to 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 aspirate the 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 aspirating the diluent.

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

[0144] The implementation of this disclosure can be described with reference to the following numbered clauses, wherein preferred features are arranged in the dependent clauses:

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

[0146] A measurement cycle is performed to 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, 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.

[0147] Calculate the first value of the first group of sampled phases;

[0148] Calculate the second value of the second set of sampled phases; and

[0149] The antibody concentration is determined based on a function of the first and second values.

[0150] 2. The method according to 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. The method according to Clause 1, wherein each sampling phase includes a voltage measurement of fluorescence from multiple light pulses emitted from the light source.

[0152] 4. The method described in Clause 3 further includes:

[0153] Multiple voltage measurements are obtained from each light pulse.

[0154] 5. The method according to 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 described in Clause 5 further includes:

[0156] Calculate the first average value of the first group of voltage measurements;

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

[0158] The light pulse difference for each light pulse is determined by subtracting the second average value from the first average value.

[0159] 7. The method described in Clause 6 further includes:

[0160] The average optical pulse difference of the sampling phase is determined by calculating the average of the optical pulse differences from the sampling phase.

[0161] 8. The method described in Clause 7 further includes:

[0162] The first polarization value is determined by calculating the average value of the average light pulse difference of the first set of sampled phases of the fluorescence emitted along the first direction.

[0163] 9. The method described in 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 sampled phases of the fluorescence emitted along the second direction.

[0165] 10. The method described in Clause 9 further includes:

[0166] The concentration value 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.

[0167] 11. The method according to Clause 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] light source;

[0170] A first polarizing filter, wherein the first polarizing filter polarizes light emitted from the light source along a first polarization direction;

[0171] A container that holds a sample mixed with a fluorescent polarizing reagent, the container receiving light polarized along the first direction;

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

[0173] Detector, the detector being used to measure 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 to measure 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 the sampling phases of the fluorescence emitted along the first direction.

[0176] Calculate the first value of the first group of sampled phases;

[0177] Calculate the second value of the second set of sampled phases: and

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

[0179] 13. The system according to Clause 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 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.

[0180] 14. The system according to Clause 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 the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0182] Multiple voltage measurements are obtained from each light 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 the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0185] Calculate the first average value of the first group of voltage measurements;

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

[0187] The light pulse difference for each light pulse is determined by subtracting the second average value from the first average value.

[0188] 18. The system according to Clause 17, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0189] The average optical pulse difference of the sampling phase is determined by calculating the average of the optical pulse differences from the sampling phase.

[0190] 19. The system according to Clause 18, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0191] The first polarization value is determined by calculating the average value of the average light pulse difference of the first set of sampled phases of the fluorescence emitted along the first direction.

[0192] 20. The system according to Clause 19, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0193] The second polarization value is determined by calculating the average value of the average light pulse difference of the second set of sampled phases of the fluorescence emitted along the second direction.

[0194] 21. The system according to Clause 20, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0195] The concentration value 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.

[0196] 22. The system according to Clause 12, wherein the antibody is immunoglobulin G (IgG).

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

[0198] Perform a measurement cycle of fluorescence emitted from the cell sample mixed with a fluorescent polarizing reagent, the measurement cycle comprising the following measurement sequence:

[0199] (1) Measure the first sampling phase of fluorescence emitted along a first direction before the midpoint of the measurement period;

[0200] (2) Measure 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) Measure the third sampling phase of the fluorescence emitted along the second direction after the midpoint of the measurement period;

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

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

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

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

[0206] 24. The method according to Clause 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 described under Clause 24 further includes:

[0208] Multiple voltage measurements are obtained from each light pulse.

[0209] 26. The method according to Clause 25, 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.

[0210] 27. The method described pursuant to Clause 26 further includes:

[0211] Calculate the average value of the first group of voltage measurements;

[0212] Calculate the average value of the second set of voltage measurements; and

[0213] 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.

[0214] 28. The method described pursuant to Clause 27 further includes:

[0215] The average voltage value for each sampling phase is determined by calculating the average of the light pulse differences among the plurality of light pulses in each sampling phase.

[0216] 29. The method described pursuant to Clause 28 further includes:

[0217] The first polarization value of the first set of measurements of the sampled phase is determined by calculating the average of the average voltage values ​​of the first set of measurements of the sampled phase.

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

[0219] The second polarization value of the second set of measurements of the sampling phase is determined by calculating the average value of the average voltage values ​​of the second set of measurements of the sampling phase.

[0220] 31. The method described under Clause 30 further includes:

[0221] The concentration value is determined by subtracting the second voltage value from the first voltage value and then dividing by the sum of the first and second voltage values.

[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 cell samples, 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) Measure the first sampling phase of fluorescence emitted along a first direction before the midpoint of the measurement period;

[0227] (2) Measure 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;

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

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

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

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

[0232] The antibody concentration is determined based on a function of the first average and the second average.

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

[0234] 35. The system according to clause 34, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0235] Multiple voltage measurements are obtained from each light pulse.

[0236] 36. The system according to Clause 35, 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.

[0237] 37. The system according to clause 36, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0238] Calculate the average value of the first group of voltage measurements;

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

[0240] 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.

[0241] 38. The system according to clause 37, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0242] The voltage value of each sampling phase is determined by calculating the average of the light pulse differences among the plurality of light pulses in each sampling phase.

[0243] 39. The system according to clause 38, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0244] The first polarization value is determined by calculating the average of the voltage values ​​of each sampling phase of the fluorescence emitted along the first direction.

[0245] 40. The system according to clause 39, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0246] The second polarization value is determined by calculating the average of the voltage values ​​of each sampling phase of the fluorescence emitted along the second direction.

[0247] 41. The system of claim 40, wherein, when the instruction is executed by the processing circuit system, the processing circuit system further causes the processing circuit system to:

[0248] The antibody concentration 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.

[0249] 42. The system according to Clause 33, wherein 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, which, when executed by the processing circuit system, cause the processing circuit system to:

[0252] A measurement cycle is performed to 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, the second direction being substantially perpendicular to the first direction. 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. Each sampling phase in the first set of sampling phases and the second set of sampling phases includes a voltage measurement of light from multiple light pulses emitted from the light source.

[0253] Multiple voltage measurements are obtained from each light pulse, wherein the multiple voltage measurements from each light pulse include 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 the average value of the first group of voltage measurements;

[0255] Calculate the average value 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 of each sampling phase is determined by calculating the average of the optical pulse differences among the plurality of optical pulses in each sampling phase;

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

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

[0260] The concentration 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 and second polarization values.

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

[0262] light source;

[0263] An excitation polarizer is used to polarize 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 holds a first detection polarizer and a second detection polarizer, the first detection polarizer restricting fluorescence emitted from the container to pass through in a first direction, and the second detection polarizer restricting fluorescence emitted from the container to pass through in a second direction perpendicular to the first direction.

[0266] Detector, the detector being used to measure 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 the rotation axis in alternating directions, such that the first detection polarizer and the second detection polarizer are alternately placed in the optical path of the fluorescence emitted from inside the container.

[0269] 45. The system according to clause 44, wherein, when the instructions are executed by the processing circuitry system, the processing circuitry system further causes the processing circuitry system to:

[0270] Rotate the polarizer holder to position the first detection polarizer in the optical path of the fluorescence emanating from within the container; and

[0271] The polarizer holder is rotated to position the second detection polarizer in the optical path of the fluorescence emanating from inside the container.

[0272] 46. ​​The system described in Clause 44 further includes:

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

[0274] The polarizer holder includes a pin that shifts within the groove of the optical plate as the polarizer holder rotates about the rotation axis.

[0275] 47. The system according to Clause 46, wherein the optical plate further comprises a first magnet positioned adjacent to the first stop in the groove and a second magnet positioned adjacent to the second stop in the groove; and

[0276] The pin is made of ferromagnetic material. When it is positioned next to the first stop at the end of the first rotation stroke, the pin is attracted to the first magnet, and when it is positioned next to the second stop at the end of the second rotation stroke, the pin is attracted to the second magnet.

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

[0278] 49. The system according to Clause 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 Clause 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 biasing element and the second biasing element bend 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 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.

[0283] 54. The system described in Clause 53 further includes:

[0284] An optical cover that at least partially houses 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 described in Clause 44 further includes:

[0286] Work platform;

[0287] A liquid storage module installed on the working platform, the liquid storage module comprising:

[0288] The base has an internal volume for accommodating an internal storage device;

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

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

[0291] A second motor, the second motor being attached to the sub-cover; and

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

[0293] When the instruction is executed by the processing circuit system, the processing circuit system also causes the processing circuit 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, the second motor is operated to close the secondary cover, thereby sealing the port on the main cover.

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

[0297] 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 restricts the emitted fluorescence to pass through in a first direction; and

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

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

[0303] 57. The polarizer holder according to Clause 56 further includes:

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

[0305] The 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 Clause 57 further includes:

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

[0308] A second biasing element is attached to the second side surface to fix the second detection polarizer in the second recess.

[0309] 59. The polarizer holder according to Clause 58, wherein the first detection polarizer and the second detection polarizer protrude beyond the first side surface and the second side surface when the first detection polarizer and the second detection polarizer are respectively placed in the first recess and the second recess, and the first biasing element and the second biasing element bend 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 retainer as described in Clause 59, wherein the first retaining spring and the second retaining spring include cutouts to increase the curvature around the first and second detection polarizers.

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

[0312] The polarizer holder is positioned in a first position such that the first detection polarizer is placed in the optical path emitting fluorescence from inside the container, which contains an antibody sample mixed with a fluorescent polarizing reagent.

[0313] After the light passes through the first detection polarizer, the fluorescence emitted from inside the container is detected, and the first detection polarizer restricts the fluorescence emitted from inside the container to pass through in a first direction;

[0314] The polarizer holder is rotated about a rotation axis to position it in a second position, such that a second detection polarizer is placed in the optical path of the 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

[0315] After the light passes through the second detection polarizer, the fluorescence emitted from inside the container is detected.

[0316] 62. The method described pursuant to Clause 61 further includes:

[0317] The polarizer holder is rotated about the rotation axis so that the polarizer holder returns 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 described pursuant to Clause 61 further includes:

[0320] The antibody concentration is determined based on the detection of light passing through the first and second detection polarizers.

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

[0322] The base has an internal volume for accommodating an internal storage device;

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

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

[0325] An electric motor, the electric motor being attached to the sub-cover; and

[0326] A processing circuit system having a memory for storing instructions, which, when executed by the processing circuit system, cause 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 reach the bottom of the internal reservoir; and

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

[0329] 66. The module described in Clause 65 further includes:

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

[0331] 67. The module described in Clause 65 further includes:

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

[0333] 68. The module according to Clause 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 motor.

[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 primary cover to keep 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 primary cover.

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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