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

Abstract

A system for measuring the concentration of antibodies in a sample includes a polarizer holder that holds a first detection polarizer and a second detection polarizer. The first detection polarizer restricts the passage of light fluorescing from within the container in a first direction. The second detection polarizer restricts the passage of light fluorescing from within the container in a second direction, which is perpendicular to the first direction. The system includes a detector for measuring light fluorescing in the first direction and light fluorescing in the second direction. The system rotates the polarizer holder around a rotation axis in alternating directions, causing the first and second detection polarizers to alternately position within the optical path of light fluorescing from within the container.

Description

[Background technology]

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

[0002] Antibodies are large, Y-shaped proteins used by the immune system to identify and neutralize foreign substances such as pathogenic bacteria and viruses. Antibodies recognize a unique molecule of a pathogen called an antigen. Each end of a Y-shaped antibody contains a paratope, which allows the antibody to bind to the antigen and match to an epitope on the antigen. Using this binding mechanism, antibodies can tag pathogens or infected cells for attack by other parts of the immune system, or they can directly neutralize pathogens or infected cells. Immunoglobulin G (IgG) is the most common type of antibody found in the blood circulation. The concentration of antibodies in cell samples can be measured by fluorescence polarization. [Overview of the project] [Means for solving the problem]

[0003] In general terms, this disclosure relates to measuring the concentration of antibodies in cell samples by fluorescence polarization. In one possible configuration, the measurement cycle is performed to reduce errors that may result from signal intensity fluctuations and noise during fluorescence polarization. Various aspects are described in this disclosure, and this includes, but is not limited to, the following aspects:

[0004] One aspect relates to a fluorescence polarization method for measuring the concentration of an antibody in a sample, comprising: performing a measurement cycle that measures a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, wherein at least one sampling phase of light that fluoresces in the second direction occurs between the sampling phases of light that fluoresces in the first direction; calculating a first value for the first set of sampling phases; calculating a second value for the second set of sampling phases; and determining the concentration of the antibody based on a function of the first and second values.

[0005] Another aspect is a fluorescence polarization system for measuring the concentration of an antibody in a sample, comprising a light source, a first polarizing filter that polarizes light emitted from the light source in a first direction, a container for holding a sample to be mixed with a fluorescence polarization assay agent, the container receiving light polarized in the first direction, a movable second and third polarizing filter, the second polarizing filter restricting the passage of fluorescent light emitted from within the container to a first direction, the third polarizing filter restricting the passage of fluorescent light emitted from within the container to a second direction, the second being substantially perpendicular to the first direction, and a detector for measuring the fluorescent light emitted in the first direction and the fluorescent light emitted in the second direction. The present invention relates to a system comprising a processing network having a memory for storing instructions, the processing network having an instruction which, when executed by the processing network, causes the processing network to perform a measurement cycle that measures a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, wherein at least one sampling phase of light that fluoresces in the second direction occurs between the sampling phases of light that fluoresces in the first direction; calculate a first value for the first set of sampling phases; calculate a second value for the second set of sampling phases; and determine the concentration of an antibody based on a function of the first and second values.

[0006] Another aspect of the method is a fluorescence polarization method for measuring the concentration of an antibody in a cell sample, the method comprising performing a measurement cycle of light fluorescing from within a cell sample mixed with a fluorescence polarization assay agent, the measurement cycle comprising measurements in the following order: (1) measuring a first sampling phase of light fluorescing in a first direction before the midpoint of the measurement cycle; (2) measuring a second sampling phase of light fluorescing in a second direction before the midpoint of the measurement cycle, the second direction being perpendicular to the first direction; (3) measuring a third sampling phase of light fluorescing in a second direction after the midpoint of the measurement cycle; and (4) measuring a fourth sampling phase of light fluorescing in a first direction after the midpoint of the measurement cycle; calculating a first average value of the first and fourth sampling phases; calculating a second average value of the second and third sampling phases; and determining the concentration of the antibody based on a function of the first and second average values.

[0007] Another aspect is a fluorescence polarization system for measuring the concentration of antibody proteins in a cell sample, the system being a processing network having a memory for storing instructions, and when an instruction is executed by the processing network, the processing network is to perform a measurement cycle of light fluorescence emitted from the cell sample, the measurement cycle being a measurement in the following order: (1) measuring a first sampling phase of light fluorescence emitted in a first direction before the midpoint of the measurement cycle, and (2) measuring a second sampling phase of light fluorescence emitted in a second direction before the midpoint of the measurement cycle. The present invention relates to a system comprising a processing network that includes (3) measuring a third sampling phase of light that fluoresces in the second direction after the midpoint of the measurement cycle, and (4) measuring a fourth sampling phase of light that fluoresces in the first direction after the midpoint of the measurement cycle, and calculating a first average value of the first and fourth sampling phases, calculating a second average value of the second and third sampling phases, and determining the concentration of the antibody based on a function of the first and second average values.

[0008] Another aspect is a fluorescence polarization system for measuring the concentration of immunoglobulin G (IgG) in a sample, wherein the system is a processing network, the processing network having a memory for storing instructions, and when an instruction is executed by the processing network, the processing network performs a measurement cycle measuring a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, the second direction being approximately perpendicular to the first direction, and at least one sampling phase of light that fluoresces in the second direction occurring between the sampling phases of light that fluoresces in the first direction, and each sampling phase in the first and second sets of sampling phases includes a voltage measurement of light from a plurality of light pulses emitted by a light source, and a plurality of voltage measurements from each light pulse, the plurality of voltage measurements from each light pulse being a first set of voltage measurements when the light source is turned on and the light source is The present invention relates to a system comprising a processing network that includes a second set of voltage measurements taken when the system is turned off, calculates the average value of the first set of voltage measurements, calculates the average value of the second set of voltage measurements, determines the optical pulse difference value for each optical pulse by subtracting the average value of the second set of voltage measurements from the average value of the first set of voltage measurements, determines the voltage value for each sampling phase by calculating the average value of the optical pulse difference values ​​of multiple optical pulses in each sampling phase, determines a first polarization value for the first set of sampling phases by calculating the average value of the voltage values ​​in the first set of sampling phases, determines a second polarization value for the second set of sampling phases by calculating the average value of the voltage values ​​in the second set of sampling phases, and determines the IgG concentration value by subtracting the second polarization value from the first polarization value and dividing by the sum of the first and second polarization values.

[0009] Another aspect is a cell analysis system for measuring the concentration of antibodies in a sample, the system comprising a light source, an excitation polarizer for polarizing light emitted from the light source in a first direction, a container configured to receive the light polarized in the first direction, and a polarizer holder for holding a first detection polarizer and a second detection polarizer, wherein the first detection polarizer restricts the passage of fluorescent light emitted from within the container to a first direction, and the second detection polarizer restricts the passage of fluorescent light emitted from within the container to a second direction, the second direction being the first direction The system comprises a polarizer holder perpendicular to the direction, a detector for measuring light that fluoresces in a first direction and light that fluoresces in a second direction, and a processing network having a memory for storing instructions, which, when executed by the processing network, causes the processing network to rotate the polarizer holder about a rotation axis in alternating directions, thereby causing the alternating placement of a first detection polarizer and a second detection polarizer in the optical path of light that fluoresces from inside the container.

[0010] Another aspect relates to 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 to the first side surface which restricts the passage of fluorescent light in a first direction, and a second detection polarizer coupled to the second side surface which restricts the passage of fluorescent light in a second direction, the polarizer holder being configured to rotate about an axis of rotation and cause alternating placement of the first and second detection polarizers in the optical path of light.

[0011] Another aspect is a fluorescence polarization method for measuring the concentration of an antibody in a sample, the method comprising: positioning a polarizer holder in a first position such that a first detection polarizer is placed in the optical path of light fluorescing from within a container holding a sample of antibody mixed with a fluorescence polarization assay agent; detecting light fluorescing from within the container after the light has passed through the first detection polarizer, wherein the first detection polarizer restricts the passage of light fluorescing from within the container in a first direction; rotating the polarizer holder about a pivot axis such that a second detection polarizer is placed in the optical path of light fluorescing from within the container, wherein the second detection polarizer restricts the passage of light fluorescing from within the container in a second direction; and detecting light fluorescing from within the container after the light has passed through the second detection polarizer.

[0012] Another aspect relates to a module for a cell analysis system, the module comprising: a base having an internal volume for holding an internal reservoir; a primary lid connected to the base, the primary lid including a port providing access to the internal reservoir; a secondary lid attached to the primary lid, the secondary lid configured to seal the port on the primary lid; a motor attached to the secondary lid; and a processing network, the processing network having a memory for storing instructions, which, when executed by the processing network, causes the processing network to operate the motor to open the secondary lid, allowing a probe to be inserted through the port on the primary lid and reach the bottom of the internal reservoir; and when the probe is removed, to operate the motor to close the secondary lid, sealing the port on the primary lid.

[0013] Various additional aspects will be described in the following explanation. This aspect can relate to individual features and combinations of features. It should be understood that both the foregoing general description and the following detailed description are merely exemplary and explanatory, and do not limit the broad inventive concept based on the embodiments disclosed herein.

Brief Description of the Drawings

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

[0015] [Figure 1] FIG. 1 is an isometric view of an embodiment of a cell analysis system for analyzing the cell viability of a plurality of cell samples.

[0016] [Figure 2] FIG. 2 is another isometric view of the cell analysis system of FIG. 1 with the upper cover removed from the housing of the cell analysis system.

[0017] [Figure 3] FIG. 3 is a top view of a work platform supported inside the housing of the cell analysis system of FIG. 1.

[0018] [Figure 4] FIG. 4 schematically illustrates an embodiment of a titer module supported on the work platform of FIG. 3.

[0019] [Figure 5] FIG. 5 graphically illustrates an embodiment of a plot showing the detected light intensity drift within the titer module of FIG. 4 that causes an error when calculating the concentration of an antibody.

[0020] [Figure 6] FIG. 6 graphically illustrates a plot showing the simulated fluorescence polarization measurement error resulting from the measurement sequence shown in the plot of FIG. 5.

[0021] [Figure 7] Figure 7 schematically illustrates one example of a method for measuring the concentration of an antibody in a cell sample, which can be carried out using the titer module shown in Figure 4.

[0022] [Figure 8] Figure 8 schematically illustrates one embodiment of a method for performing a measurement cycle in the operation of the method shown in Figure 7.

[0023] [Figure 9] Figure 9 schematically illustrates an embodiment of a measurement cycle carried out according to the method in Figure 8.

[0024] [Figure 10] Figure 10 graphically illustrates an example with plots showing the extent to which the method in Figure 7 reduces errors when calculating antibody concentrations in cell samples.

[0025] [Figure 11] Figure 11 graphically illustrates the plots showing the reduced fluorescence polarization measurement error resulting from the measurement cycles in Figures 8-10.

[0026] [Figure 12] Figure 12 schematically illustrates the method for measuring the sampling phase in the measurement cycle shown in Figures 8-10.

[0027] [Figure 13] Figure 13 schematically illustrates one embodiment of the controller of the cell analysis system of Figure 1, which may be used to implement the side view of the titer module of Figure 4.

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

[0029] [Figure 15] Figure 15 is an isometric view of the titer module from Figure 14, with part of the casing removed to expose the internal components of the titer module.

[0030] [Figure 16] Figure 16 is an isometric cross-sectional view obtained along the horizontal axis of the titer module in Figure 14.

[0031] [Figure 17] Figure 17 is an isometric view of the titer module from Figure 14, with the internal cover removed and the optical components of the titer module exposed.

[0032] [Figure 18] Figure 18 is a right-side view of the optical components of the titer module shown in Figure 14.

[0033] [Figure 19] Figure 19 is a left side view of the optical components of the titer module shown in Figure 14.

[0034] [Figure 20] Figure 20 is an isometric cross-sectional view obtained along the vertical axis of the titer module in Figure 14.

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

[0036] [Figure 22] Figure 22 is a left cross-sectional view of the titer module shown in Figure 14.

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

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

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

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

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

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

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

[0044] [Figure 30] Figure 30 is an isometric view of the liquid storage module of the cell analysis system shown in Figure 1.

[0045] [Figure 31] Figure 31 is another isometric view of the liquid storage module from Figure 30, showing the lid of the liquid storage module in the open position.

[0046] [Figure 32] Figure 32 is another isometric view of the liquid storage module from Figure 30, showing the internal reservoir partially removed from the inside of the liquid storage module.

[0047] [Figure 33]Figure 33 is another isometric view of the liquid storage module from Figure 30, showing the automated lid of the liquid storage module in the open position.

[0048] [Figure 34] Figure 34 is another isometric view of the liquid storage module from Figure 30, showing the lid of the liquid storage module in the open position.

[0049] [Figure 35] Figure 35 is an isometric view of the secondary lid of the liquid storage module shown in Figure 30.

[0050] [Figure 36] Figure 36 is a front view of the secondary cover shown in Figure 35.

[0051] [Figure 37] Figure 37 is a side view of the secondary cover shown in Figure 35.

[0052] [Figure 38] Figure 38 is a bottom view of the secondary lid shown in Figure 35.

[0053] [Figure 39] Figure 39 is an isometric view of the inside of the cell analysis system in Figure 1, showing the dispensing system that positions the probe near the liquid storage module in Figure 30.

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

[0055] [Figure 41] Figure 41 illustrates an example of an alternative embodiment of the polarizer holder for the titer module shown in Figure 14. [Modes for carrying out the invention]

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

[0057] The cell analysis system 100 includes connectivity to an automated bioreactor and other systems and devices. This connectivity may include electrical connectivity for data sharing and physical connectivity for liquid handling purposes. The cell analysis system 100 automates sample preparation and minimizes sample volume requirements for measuring cell properties. It provides remote access to data, including measured cell properties, supports multiple users simultaneously, and is compatible with various information technology (IT) structures.

[0058] As shown in Figure 1, the cell analysis system 100 includes a housing 102 that supports a work platform 300. The housing 102 includes a top cover 104, which may support a dispensing system 106, which will be described in more detail with reference to Figures 2 and 3.

[0059] Figure 2 is another isometric view of the cell analysis system 100, with the top cover 104 removed from the housing 102, thereby exposing the dispensing system 106. Figure 3 is a top view of the work platform 300 supported inside the housing 102. Referring here to Figures 2 and 3, the work platform 300 supports one or more tube racks that hold multiple containers containing cell samples and various types of reagents. In some cases, at least some of the containers are empty. The dispensing system 106 includes a probe 108 that is movably mounted inside the housing 102 in the space above the work platform 300. The dispensing system 106 is one embodiment 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). Three-dimensional movement allows the distal end of the probe 108 (through which liquid aspiration and dispensing occur) to access any container held on the work platform 300 inside the housing 102.

[0061] The proximal end of the probe 108 is fluidly connected to a bidirectional pump having a movable actuator that controls the pump's mode. For example, a first mode may include aspiration, and a second mode may include dispensing, with the movable actuator controlling the switching between the first and second modes and the rates of fluid aspiration and dispensing under the first and second modes. In embodiments, the bidirectional pump may include a syringe pump. The movement of the probe 108 and the movable actuator is controlled by one or more stepper motors operating under the control of a programmable controller 1300.

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

[0063] As shown in Figure 3, the work platform 300 supports a sample transfer module 302, one or more mixed plates 304, a cell health module 306, one or more tip racks 308, one or more titer plates 310, a tip waste container 312, a liquid storage module 314, sample inputs 316 such as tube trays and well plates, a metabolite module 318, and a titer module 400. The cell health module 306 measures cell health, which includes cell number and cell viability. The titer module 400 measures protein titer, which will be described in more detail here with reference to Figure 4.

[0064] Figure 4 schematically illustrates an embodiment of the titer module 400 supported on a work platform 300. The titer module 400 includes optical components used to measure 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 determine the concentration of IgG protein in a cell sample. Fluorescence polarization involves mixing the cell sample with a fluorescence polarization assay agent 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 in reference to measuring IgG concentration, the titer module 400 and the measurement techniques described herein can be used to measure the concentrations of additional types of proteins and antibodies.

[0066] As shown in Figure 4, the titer module 400 includes a light source 402 that emits light 404 toward a container 412 holding a solution of cells to be mixed with a fluorescence polarizing assay agent. The fluorescence polarizing assay agent is conjugated with an antibody of interest (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 fluorescence polarizing assay agent and the sample of cells in the container 412 together, so that the user of the system does not need to manually mix the solution in the container 412. In some embodiments, the container 412 is a cuvette, tube, and equivalent.

[0067] The light source 402 emits unpolarized light 404 such that the light 404 is unpolarized light. In some embodiments, the light source 402 is a light-emitting diode (LED). In some embodiments, the titer module 400 includes a focusing lens 406 that focuses the light 404 emitted from the light source 402 and a spectral filter 408 that filters the light 404 emitted from the light source 402.

[0068] Light 404 passes through an excitation polarizer 410 that polarizes the light 404 in a first direction 411. In some embodiments, the first direction 411 is linear. In the embodiment shown in Figure 4, the first direction 411 is vertically linear. In alternative embodiments, the first direction 411 is horizontally linear. Additional polarization directions are also possible.

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

[0070] The polarized fluorescent light 416 can pass through the lens 414 for focusing after being emitted from the solution in the container 412. The polarized fluorescent light 416 then alternately passes through the first detection polarizer 420 and the second detection polarizer 422. The first detection polarizer 420 and the second detection polarizer 422 are mounted in a polarizer holder 418, which is controlled by the controller 1300 to cause the alternating placement of the first detection polarizer 420 and the second detection polarizer 422 in the path of the polarized fluorescent light 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 restricts the passage of the polarized fluorescent light 416 to the first direction 411.

[0072] The second detection polarizer 422 is polarized in a second direction 413, which is perpendicular to the first direction 411 of the excitation polarizer 410. The second detection polarizer 422 restricts the passage of the polarized fluorescent light 416 to the second direction 413. The polarized fluorescent light 416 then passes through the spectral filter 424 before detection by the detector 426. In some embodiments, the detector 426 is a photomultiplier tube (PMT). The detector 426 converts the polarized fluorescent light 416 into a voltage value for input to equation 1 in order to determine the concentration of antibodies produced by identical cells.

number

[0073] The measured fluorescence polarization (FP) is correlated with the concentration of antibody in a sample of cells. For example, a higher detected FP correlates with a higher concentration of antibody, and a lower detected FP correlates with a lower concentration of antibody. This is because a higher concentration of antibody has an increased binding with the fluorescence polarization assay agent, and as a result, the voltage of the parallel fluorescence becomes larger and the voltage of the perpendicular fluorescence becomes smaller, which results in a larger detected FP.

[0074] The technical difficulty of Equation 1 is that any error in measuring I PER that does not exist within I PAR can cause an error in the calculation of fluorescence polarization (FP). For example, the FP calculation depends on the measured ratio I [[ID=⑨]]<00,000,⑦> / I PAR . Any fluctuations that affect I PAR and I PER proportionally are canceled out. However, error such as signal intensity fluctuations throughout the process of measuring parallel fluorescence (I PAR ) and perpendicular fluorescence (I PER ), as well as offset errors that affect both perpendicular fluorescence (I PER ) and parallel fluorescence (I PAR ) are not canceled out and cause significant errors in the FP calculation, which can thereby cause errors when calculating the concentration of antibody in a sample of cells.

[0075] Sources of signal intensity fluctuations can include, but are not limited to, increases and decreases in the intensity and drift of the light source 402, electrical response, dye bleaching, and non-uniform fluid and diffusion within the container 412. Additional sources of error are also possible. Sources of offset noise can include, but are not limited to, electrical circuit noise and ambient light.

[0076] [[ID=3①]] Figure 5 graphically illustrates an example with plot 500 showing the light intensity drift detected in the titer module 400, which causes errors in FP calculations and miscalculations of antibody concentrations in cell samples. Plot 500 includes light intensity (Y axis) measured by detector 426 over time (X axis). In this example, light intensity is measured without using any solution of cells or fluorescence polarizing assay reagents in container 412 so that light intensity drift is not caused by the solution.

[0077] In this embodiment, the light intensity of the light source 402 is expected to have a constant value of 3.5V. Therefore, PER and I PAR The ratio between fluorescence should be 1:1 (e.g., 3.5V:3.5V) because there are no antibody-containing cell samples in container 412.

[0078] However, the actual light intensity shown in plot 500 is I PER and I PAR It exhibits voltage drift that causes errors in the ratio between fluorescence. In this example, I PER Fluorescence is measured in the first phase (i.e., using the second detection polarizer 422), and then I PAR A measurement cycle is performed in which fluorescence is measured in the second phase (i.e., using the first detection polarizer 420). PER and I PAR Each fluorescence phase includes two measurements. For example, I PER The fluorescence includes a first measurement at 3.60V and a second measurement at 3.62V, I PAR Fluorescence includes a third measurement at 3.64V and a fourth measurement at 3.66V. PER Fluorescence is calculated as the average of the first and second measurements (i.e., 3.61V), I PAR Fluorescence is calculated as the average of the third and fourth measurements (i.e., 3.65V). This is I PER and I PARThis results in a 3.61:3.65 ratio between fluorescence, which is an error of approximately 1% caused by the drift of light source 402.

[0079] Figure 6 graphically illustrates plot 600, showing the simulated fluorescence polarization measurement error resulting from the measurement sequence in Figure 5. As shown in Figure 6, the measurement error is significantly larger for lower mP values, such as mP values ​​of less than 200 units. Therefore, lower concentrations of antibody in the sample held by the container are particularly sensitive to signal intensity fluctuations and noise in the titer module 400.

[0080] In some cases, I2 is affected by signal strength fluctuations and noise. PER and I PAR Measurement errors in the ratio between fluoresces can be reduced by implementing complex electrical circuit designs and temperature-controlled light sources and detectors within the titer module. However, such solutions are expensive, unreliable, increase the size of the titer module, and can slow down the preheating time required to use the device.

[0081] Figure 7 schematically illustrates one embodiment of method 700 for measuring the concentration of antibodies in a cell sample. Method 700 uses the optical components of the titer module 400 (shown in Figure 4) to calculate the concentration of antibodies in a cell sample. In some embodiments, method 700 is performed to measure the concentration of immunoglobulin G (IgG) in a solution containing a fluorescence polarizing assay agent mixed in a container 412, using the dispensing system 106 of the cell analysis system 100.

[0082] Method 700 reduces errors from signal intensity fluctuations and noise without requiring any changes to the optical components and hardware of the titer module 400. Instead, Method 700 is a software solution that improves the functionality of the titer module 400 by reducing these types of errors to improve the accuracy of antibody concentration calculations.

[0083] Method 700 includes an operation 702 for performing a measurement cycle. Operation 702 includes using the optical components of the titer module 400 to perform a first set of measurements of light emitted from the solution in a first direction 411, and a second set of measurements of light emitted from the solution in a second direction 413. The first and second sets of measurements alternate with each other in the measurement cycle.

[0084] Figure 8 schematically illustrates an embodiment of Method 800 in which a measurement cycle is performed in operation 702 of Method 700. Figure 9 schematically illustrates an embodiment of a measurement cycle 900 performed according to Method 800.

[0085] Referring here to Figures 8 and 9, Method 800 includes step 802 of measuring a first sampling phase A of light fluorescently emitted from the container 412 in a first direction 411, before the midpoint 902 of the measurement cycle 900. Next, Method 800 includes step 804 of measuring a second sampling phase B of light fluorescently emitted from the container 412 in a second direction 413, before the midpoint 902 of the measurement cycle 900.

[0086] As further shown in Figures 8 and 9, Method 800 includes step 806 of measuring a third sampling phase C of light fluorescently emitted from the container 412 in a second direction 413 after the midpoint 902 of the measurement cycle 900. Next, Method 800 includes step 808 of measuring a fourth sampling phase D of light fluorescently emitted from the container 412 in a first direction 411 after the midpoint 902 of the measurement cycle 900.

[0087] The measurement cycle 900 includes at least one sampling phase in a first direction 411 before the midpoint and at least one sampling phase in a second direction 413. The measurement cycle includes at least one sampling phase in a first direction 411 after the midpoint of the measurement cycle and at least one sampling phase in a second direction 413.

[0088] During the measurement cycle 900, the polarizer holder 418 is controlled by the controller 1300 to switch at transition point 904 from the first detection polarizer 420, which is polarized in a direction parallel to the direction of the excitation polarizer 410, to the second detection polarizer 422, which is polarized in a direction perpendicular to the direction of the excitation polarizer 410. Transition point 904 occurs between the first and second sampling phases A and B. Also during the measurement cycle 900, the polarizer holder 418 is controlled by the controller 1300 to switch at transition point 906 from the second detection polarizer 422 (polarized in a direction perpendicular to the direction of the excitation polarizer 410) to the first detection polarizer 420 (polarized in a direction parallel to the direction of the excitation polarizer 410). Transition point 906 occurs between the third and fourth sampling phases C and D.

[0089] As further shown in Figure 9, each sampling phase A and D includes voltage measurements of fluorescent light emitted from multiple light pulses 908 emitted by the light source 402. For each light pulse 908, the detector 426 acquires multiple voltage measurements 910. For example, the multiple voltage measurements 910 from each light pulse 908 include a first set 910a of voltage measurements when the light source 402 is turned on and a second set 910b of voltage measurements when the light source 402 is turned off. Steps 802-808 for measuring the first, second, third, and fourth sampling phases A, B, C, and D may each include steps to perform additional operations as shown in Figure 12.

[0090] Figure 12 schematically illustrates a method 1200 for measuring the sampling phase, as performed in the measurement cycle of Figures 8-10. Referring here to Figures 9 and 12, method 1200 includes operation 1202 for calculating the average value of a first set 910a of voltage measurements when the light source 402 is turned on. Next, method 1200 includes operation 1204 for calculating the average value of a second set 910b of voltage measurements when the light source 402 is turned off. Next, method 1200 includes operation 1206 for determining the voltage value for each optical pulse 908 in the sampling phase by subtracting the average value of the second set 910b of voltage measurements from the average value of the first set 910a of voltage measurements. Next, method 1200 may include operation 1208 for determining the voltage value for the sampling phase by calculating the average value of the voltage values ​​of a plurality of optical pulses 908 in the sampling phase.

[0091] Referring back to Figure 7, method 700 includes operation 704 for calculating the first average value of the first and fourth sampling phases A and D. As shown in Figure 9, the first sampling phase A includes light that fluoresces in the first direction 411 before the midpoint 902 of the measurement cycle 900. The fourth sampling phase D includes light that fluoresces in the first direction 411 after the midpoint 902 of the measurement cycle 900.

[0092] Next, method 700 includes operation 706 for calculating the second average value of the second and third sampling phases B and C. As described above, the second sampling phase B includes light emitted from the container in the second direction 413 before the midpoint 902 of the measurement cycle 900. The third sampling phase C includes light emitted from the container in the second direction 413 after the midpoint 902 of the measurement cycle 900.

[0093] Method 700 includes operation 708, which determines the concentration of the antibody based on the ratio of a first and a second mean value. For example, the first mean value is parallel fluorescence (I PAR The voltage of ) and the second average value is right-angle fluorescence (IPER The voltage is ), and the antibody concentration is calculated using equation 1.

[0094] Figure 10 graphically illustrates an example with plot 1000, showing the extent to which method 700 eliminates errors in calculating the units (mP) of polarization measurements used to determine the concentration of antibodies in a sample of cells. Plot 1000 includes light intensity (Y axis) measured by detector 426 over time (X axis). As shown in the exemplary plot in Figure 5, plot 1000 shows light source 402 exhibiting a drift from an expected value of 3.5 V. Light intensity is measured without using any solution of cells and fluorescence polarization assay reagents in container 412 so that the drift of light intensity is not caused by the solution.

[0095] As shown in Figure 10, a measurement cycle is performed in which the first sampling phase A is measured with respect to the light that fluoresces in the first direction before the midpoint M (3.55V), the second sampling phase B is measured with respect to the light that fluoresces in the second direction before the midpoint M (3.59V), the third sampling phase C is measured with respect to the light that fluoresces in the second direction after the midpoint M (3.61V), and the fourth sampling phase D is measured with respect to the light that fluoresces in the first direction after the midpoint M (3.65V). The first average of the first and fourth sampling phases A and D is calculated as 3.6V. The second average of the second and third sampling phases B and C is calculated as 3.6V. PER and I PAR This results in a 3.6:3.6 ratio between fluorescence, which eliminates errors caused by the drift of light source 402 (see plot 500 shown in Figure 5 for comparison). Thus, the measurement cycle performed in method 800 reduces errors when measuring antibody concentrations in cell samples using fluorescence polarization, and in some cases, eliminates errors altogether.

[0096] Figure 11 graphically illustrates plot 1100, which shows the reduction in simulated fluorescence polarization measurement error resulting from the measurement cycle shown in Figures 8-10. Referring here to Figures 6 and 11, plot 1100 shows that the measurement error is significantly less for lower mP values ​​(e.g., mP values ​​less than 200 units) than the measurement error shown in plot 600 when the conventional measurement sequence is performed. Plot 1100 shows that the sensitivity to signal intensity fluctuations and noise in the titer module 400 is significantly reduced for lower antibody concentrations.

[0097] Figure 13 schematically illustrates one embodiment of a controller 1300 of a cell analysis system 100 that may be used to implement aspects described herein, including features of the titer module 400. As shown in Figure 13, the controller 1300 includes one or more processing devices 1302, a memory storage device 1304, and a system bus 1306 that connects the memory storage device 1304 to one or more processing devices 1302. One or more processing devices 1302 may include a central processing unit (CPU). In some cases, one or more processing devices 1302 are part of a processing network having a memory for storing instructions that, when executed by the processing network, cause the processing network to implement various aspects, features, and functionalities described herein.

[0098] As shown in Figure 13, the memory storage device 1304 may include a random access memory ("RAM") 1308 and a read-only memory ("ROM") 1310. Basic input and output logic, which has basic routines useful for transferring information between elements within the controller 1300 during startup, etc., may be stored in the ROM 1310.

[0099] The controller 1300 may also include a mass storage device 1312, which includes an operating system 1314 and can 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 its associated computer-readable data storage medium provide a non-volatile, non-transient storage device for the controller 1300.

[0100] The description of computer-readable data storage media contained herein refers to the mass storage device 1312, but it should be understood by those skilled in the art that the computer-readable data storage medium may be any available non-transient physical device or product from which the controller 1300 can read data and / or instructions. The computer-readable storage medium may consist of a completely non-transient medium. The mass storage device 1312 is one embodiment of a computer-readable storage device.

[0101] Computer-readable data storage media include volatile and non-volatile removable and non-removable media implemented in any way or technique for storing information such as computer-readable software instructions, data structures, program modules, or other data. Exemplary types of computer-readable data storage media include, but are not limited to, RAM, ROM, EPROM, EEPROM, flash memory, or other solid-state memory technologies, or any other media that may be used to store and accessed by this device.

[0102] The controller 1300 can operate in a networked environment using logical connections to other devices through the network 1320. The controller 1300 connects to the network 1320 through a network interface unit 1318 connected to the system bus 1306. The network interface unit 1318 can also connect to additional types of communication networks and devices, including Bluetooth®, Wi-Fi, and cellular telecommunications networks, including 4G and 5G networks. The network interface unit 1318 can connect the controller 1300 to additional networks, systems, and devices. The controller 1300 also includes an input / output unit 1322 for receiving inputs from peripheral devices and processing outputs to them.

[0103] The mass storage device 1312 and RAM 1308 can store software instructions and data. The software instructions may include an operating system 1314 which is suitable for controlling the operation of the cell analysis system 100. The mass storage device 1312 and / or RAM 1308 can also store software instructions and data 1216 which, when executed by the processing device 1302, provide the functionality of the cell analysis system 100 discussed herein.

[0104] Figure 14 is an isometric view of one embodiment of a titer module 400 that may be used in a cell analysis system 100 (see also, for example, Figures 3 and 4). Figure 15 is an isometric view of the titer module 400 with a portion of the housing 401 removed, exposing the internal components of the titer module. Figure 16 is an isometric cross-sectional view obtained along the horizontal axis of the titer module 400. Referring here to Figures 14-16, the internal components of the titer module 400 include an excitation assembly 403 and a detection assembly 405.

[0105] The excitation assembly 403 includes, at least in part, a first optical cover 407 that houses the light source 402 and the excitation polarizer 410. The detection assembly 405 includes, at least in part, a detector 426, a polarizer holder 418, and a second optical cover 409 that houses the first detection polarizer 420 and the second detection polarizer 422.

[0106] 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 so that they are parallel and perpendicular to the excitation polarization axis of the excitation polarizer 410, respectively. This arrangement is generally more precise than those in which the polarizer axes are rotated, which can generally result in higher variability of polarizers in the optical path. The polarizer holder 418 addresses the technical problem of reproducible and precise switching between the first detection polarizer 420 and the second detection polarizer 422 on the detection section of the optical path within the titer module 400.

[0107] Figure 17 is an isometric view of the titer module 400 with the first and second optical covers 407, 409 removed, exposing the optical components of the titer module. Figure 18 is a right side view of the optical components of the titer module 400. Figure 19 is a left side view of the optical components of the titer module 400. The optical components of the titer module 400 shown in Figures 17-19 are substantially similar to the optical components schematically shown in Figure 4. For example, the titer module 400 includes a light source 402 that emits light toward a container 412 holding a cell culture of IgG protein to be mixed with a fluorescence polarizing assay agent. In some embodiments, the container 412 is a cuvette, tube, and equivalent.

[0108] The light source 402 emits unpolarized light such that the light is unpolarized light. In some embodiments, the light source 402 is a light-emitting diode (LED). In some embodiments, the titer module 400 includes an optical holder 460, which includes a focusing lens 406 for focusing the light emitted from the light source 402 and a spectral filter 408 for filtering the light emitted from the source 402.

[0109] Light passes through an excitation polarizer 410 that polarizes the light in a first direction 411 (see Figure 4). Once polarized in the first direction 411, the light is absorbed by a cell solution that is mixed with a fluorescence polarizing assay agent in a container 412. This causes the solution in the container 412 to emit polarized fluorescence.

[0110] The polarized fluorescent light, 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 on polarizer holder 418. As will be described in more detail, polarizer holder 418 is powered by motor 428 to rotate, which causes the alternating arrangement of the first detection polarizer 420 and the second detection polarizer 422 in the path of the polarized fluorescent light emitted from the solution in container 412.

[0111] As discussed 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.

[0112] The polarized fluorescent light then passes through the spectral filter 424 before reaching the detector 426. In some embodiments, the detector 426 is a photomultiplier tube (PMT). As described above, the detector 426 converts the polarized fluorescent light into a pressure value for input to equation 1 in order to determine the concentration of antibodies produced by the cell culture.

[0113] Figure 20 is an isometric cross-sectional view obtained along the vertical axis of the titer module 400. Figure 21 is a cross-sectional right side view of the titer module 400. Figure 22 is a cross-sectional left side view of the titer module 400. Referring here to Figures 20-22, the polarizer holder 418 is connected to the drive shaft 434 of the motor 428. In some embodiments, the motor 428 is a stepper motor or a similar type of electric motor (also known as a stepper motor or stepping motor). In other embodiments, the motor 428 is a solenoid. The motor 428 is mounted on the underside of the optical plate 419 that supports the optical components of the titer module 400.

[0114] The controller 1300 controls the motor 428 to rotate the polarizer holder 418 clockwise in direction D1 and counterclockwise in direction D2 about the rotation axis RR (see Figure 27), which causes the alternating placement of the first detection polarizer 420 and the second detection polarizer 422 in the optical path of the polarized fluorescent light emitted from the solution in the container 412 before the light reaches the detector 426. In some embodiments, the controller 1300 controls the motor 428 to rotate the polarizer holder 418 about 90 degrees clockwise in direction D1 and about 90 degrees counterclockwise in direction D2 about the rotation axis RR.

[0115] In an alternative embodiment, Figure 41 illustrates an embodiment of a polarizer holder 418a that includes a rotating mount 462 and an electromagnetic drive 464, which can be used to position a detection polarizer 466 in the path of light emitted fluorescently from a container 412. In some embodiments, the electromagnetic drive 464 drives the rotating mount 462 to rotate the detection polarizer 466 about an axis parallel to the light beam incident on the detection polarizer. In this alternative embodiment, the electromagnetic drive 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 then 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 embodiment, the electromagnetic drive 464 can rotate the detection polarizer by about 90 degrees between the first and second positions. In this embodiment, the electromagnetic drive unit 464 includes a magnet 468 positioned next to the rotating mount unit 462, which drives the rotating mount unit 462 and rotates the detection polarizer 466 to a desired rotational position. In some embodiments, the magnet 468 surrounds, or at least partially surrounds, the rotating mount unit 462 and the detection polarizer 466. The electromagnetic drive unit 464 can provide greater accuracy than a rack-and-pinion mechanism used to position the detection polarizer.

[0116] Figures 23 and 24 are isometric views of the polarizer holder 418, optical plate 419, and detector 426 of the titer module 400. Figures 25 and 26 are exploded views of the polarizer holder 418. Figure 27 is a top view of the polarizer holder 418. Referring here to Figures 23-27, 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 embodiments, the first side surface 430 and the second side surface 432 are perpendicular to each other. The polarizer holder 418 further 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 and the second side surface 432 and the curved surface 431.

[0117] The first detection polarizer 420 is mounted on the first side surface 430 of the polarizer holder 418, and the second detection polarizer 422 is mounted on the second side surface 432 of the polarizer holder 418. For example, the first side surface 430 includes a recessed portion 435, within which the first detection polarizer 420 is installed. Similarly, the second side surface 432 includes a recessed portion 435, within which the second detection polarizer 422 is installed. As shown in Figure 27, when the first detection polarizer 420 and the second detection polarizer 422 are installed inside the recessed portion 435, they protrude beyond the individual first side surface 430 and second side surface 432.

[0118] As shown in Figure 25-27, the biasing element 436 is used to hold the first detection polarizer 420 inside the recessed portion 435 of the first side surface 430, and the second detection polarizer 422 inside the recessed portion 435 of the second side surface 432. Fasteners 438, such as screws, are screwed through openings 440 on the biasing element 436 into holes 441 on the first side surface 430 and the second side surface 432, thereby attaching the biasing element 436 to the first side surface 430 and the second side surface 432, and thereby fixing the first detection polarizer 420 and the second detection polarizer 422 within the recessed portion 435. In some embodiments, the biasing element 436 is a retaining spring.

[0119] The biasing elements 436 are flexible so that they flex around the edges of the first and second detection polarizers 420 and 422, fixing the first and second detection polarizers 420 and 422 in place. In some embodiments, the biasing elements 436 include cutouts 439 for increasing their flexure. In some embodiments, the cutouts 439 are provided toward the top and bottom of each biasing element 436 to increase the flexure of the biasing elements 436 around the top and bottom of the first and second detection polarizers 420 and 422.

[0120] The biasing element 436 includes an aperture 442, which allows polarized fluorescent light emitted from the solution in the container 412 to pass through the first detection polarizer 420 and the second detection polarizer 422. Each recessed portion 435 includes an aperture 437 (see Figure 25), which aligns with a separate aperture 444 on the curved surface 431 (see Figure 26), allowing polarized fluorescent light to pass through the polarizer holder 418 to reach the detector 426.

[0121] As shown in Figure 16, the curved surface 431 of the polarizer holder 418 aligns with the curved wall 417 of the second optical cover 409, minimizing the space between the polarizer holder 418 and the curved wall 417. This can help reduce the amount of stray light that could interfere with the voltage value detected by the detector 426 entering the first and second detection polarizers 420 and 422 and reaching the detector 426. Thus, the curved surface 431 of the polarizer holder 418 can reduce noise in the voltage value detected by the detector 426.

[0122] Figure 28 is an isometric view of the optical plate 419 and motor 428 of the titer module 400 from which the polarizer holder has been removed. Figure 29 is a top view of the optical plate 419. Referring now to Figures 28 and 29, as discussed above, the polarizer holder 418 is fixed directly 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 dowel pin 446 on the underside of the holder. The dowel pin 446 slides inside the groove 448 of the optical plate 419 as the polarizer holder 418 rotates clockwise in D1 and counterclockwise in D2 about the axis of rotation RR (see Figures 20-22 and 27). As shown in Figures 28 and 29, the groove 448 is curved along the axis of rotation RR.

[0123] The dowel pin 446 contacts a first stop 450 and a second stop 451 in the groove 448 at the end of its rotation in the clockwise direction D1 and the counterclockwise direction D2, respectively. For example, when the polarizer holder 418 rotates 90 degrees in the clockwise direction D1 to position the first detection polarizer 420 in the optical path of light, the dowel pin 446 contacts the first stop 450 at the distal end of the groove 448, preventing further rotation of the polarizer holder 418. Similarly, when the polarizer holder 418 rotates 90 degrees in the counterclockwise direction D2 to position the second detection polarizer 422 in the optical path of light, the dowel pin 446 contacts the second stop 451 at the opposite distal end of the groove 448, preventing further rotation of the polarizer holder 418.

[0124] The first stop 450 and the second stop 451 are associated with the positions of the polarizer holder 418, respectively. For example, the first stop 450 is associated with the parallel polarizer position 454 when the first detection polarizer 420 is positioned in the optical path of light (see Figure 27). The second stop 451 is associated with the perpendicular position 456 when the second detection polarizer 422 is positioned in the optical path of light.

[0125] As further shown in Figures 28 and 29, the optical plate 419 includes magnets 452 positioned next to the first stop 450 and the second stop 451, respectively. Dowel pins 446 are made of a ferromagnetic material such as steel, such that they attract the dowel pins 446 when the magnets 452 are positioned close to the first stop 450 at the end of their rotational progression in the clockwise direction D1, and when the magnets 452 are positioned close to the second stop 451 at the end of their rotational progression in the counterclockwise direction D2.

[0126] The controller 1300 is programmed to cut off power from the motor 428 at the end of rotation in clockwise and counterclockwise directions D1 and D2, so that the magnet 452 firmly pulls the dowel pin 446 upward relative to 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 right-angle polarizer position, improving the overall accuracy and repeatability of the titer module 400.

[0127] The arrangement of polarizer holders 418, which rotate in clockwise direction D1 and counterclockwise direction D2 to alternately position the first detection polarizer 420 and the second detection polarizer 422 within the optical path of light emitted from the solution in container 412, reduces the space required for the polarizer holders 418 on the optical plate 419 of the titer module 400. The arrangement of polarizer holders 418 also reduces the amount of 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 with respect to the excitation polarization axis from the excitation assembly 403 of the titer module 400.

[0128] Figure 30 is an isometric view of the liquid storage module 314 of the cell analysis system 100 (see also Figure 3). In Figure 30, the liquid storage module 314 is shown in the closed position. The liquid storage module 314 can be used in combination with the titer module 400. For example, the liquid storage module 314 can store liquids such as diluents used by the titer module 400. The liquid storage module 314 is designed to enable efficient loading and pipetting of liquid diluents while preventing vaporization of the liquid diluents (especially liquid diluents that are sensitive to exposure to light and air). In an illustrative example, the liquid storage module 314 can limit vaporization to less than approximately 1.5% by volume over its onboard storage life of approximately 31 days inside the cell analysis system 100.

[0129] Figures 31 and 32 are isometric views of the liquid storage module 314 in the open position. Referring here to Figures 30-32, the liquid storage module 314 includes a base 3002 having an internal volume 3004 for holding an internal reservoir 3006. The user can access the internal reservoir 3006 (for example, to load liquid diluent into the internal reservoir 3006) by manually opening the primary lid 3008. The internal reservoir 3006 is reusable and replaceable. For example, the internal reservoir 3006 can be removed from the internal volume 3004 for cleaning or replacement with another internal reservoir (for example, a new or clean one).

[0130] In some embodiments, the internal reservoir 3006 contains a single well with a volume of approximately 40 mL. In some embodiments, the internal reservoir 3006 has a V-shaped bottom. Once the diluent is loaded into the internal reservoir 3006, the user can manually close the primary lid 3008. The primary lid 3008 remains closed during the operation of the cell analysis system 100.

[0131] The primary lid 3008 is manually pivoted around a hinge 3012 and can move between a closed position (Figure 30) and an open position (Figures 31 and 32). The liquid storage module 314 includes a gasket 3014 surrounding the internal volume 3004 in which the internal reservoir 3006 is held. The gasket 3014 can be made from ethylene propylene diene monomer (EPDM) rubber. When the primary lid 3008 is in the closed position, the gasket 3014 engages with the internal surface 3016 of the primary lid 3008, sealing around the internal reservoir 3006.

[0132] The primary 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. The attraction of one or more first magnets 3018 to one or more second magnets 3020 allows the primary cover 3008 to be held closed when the cover is pivoted to the closed position around the hinge 3012.

[0133] Figure 33 is another isometric view of the liquid storage module 314, showing the primary lid 3008 in the closed position. Referring here to Figures 30 and 33, the liquid storage module 314 includes a secondary lid 3010 that attaches to the primary lid 3008. The secondary lid 3010 is controlled by the controller 1300 to open and close as needed for the probe 108 of the dispensing system 106 (see Figure 2) to access the internal reservoir 3006. For example, the controller 1300 opens the secondary lid 3010 during a pipetting operation and closes it after the pipetting operation is complete, sealing the diluent inside the internal reservoir 3006, thereby preventing the diluent from vaporizing during use of the cell analysis system 100.

[0134] As shown in Figure 33, when the secondary lid 3010 is opened, the port 3022 on the primary lid 3008 is exposed. The probe 108 of the dispensing system 106 can be inserted through the port 3022 to access the internal reservoir 3006 without having to open the primary lid 3008. In some embodiments, the port 3022 is sized to allow a 55 μL or 230 μL tapered pipette tip to pass through the port. Such pipettes are typically not viable for use with solid diaphragm caps.

[0135] The secondary cover 3010 has a gasket 3024 to seal the port 3022 and prevent vaporization of the diluent contained inside the internal reservoir 3006 when the secondary cover 3010 is in the closed position. In some embodiments, the gasket 3024 is made from polyethylene (PE) foam.

[0136] Figure 34 is another isometric view of the liquid storage module 314 showing the primary lid 3008 in the open position. Referring here to Figure 34, a magnet 3026 is mounted on the primary lid 3008, which attracts an iron element 3028 on the secondary lid 3010, thereby closing the secondary lid 3010. For example, the hinge 3030 of the secondary lid 3010 may include an iron element 3028 that is attracted to the magnet 3026 and closes the secondary lid 3010. In an alternative embodiment, the magnet 3026 is located on the secondary lid 3010 and the iron element 3028 is located on the primary lid 3008. The magnet 3026 and iron element 3028 keep the secondary lid 3010 closed.

[0137] Motor 3032 is mounted on the inner surface 3016 of the primary cover 3008. The secondary 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 the secondary cover 3010 between an open and closed state. Motor 3032 can track the open and closed states of the secondary cover 3010 using an integrated encoder.

[0138] Figure 35 is an isometric view of the secondary cover 3010. Figure 36 is a front view of the secondary cover 3010. Figure 37 is a side view of the secondary cover 3010. Figure 38 is a bottom view of the secondary cover 3010. Referring now to Figures 35-38, the secondary cover 3010 includes an upper portion 3040 and a hinge 3042 extending from the upper portion 3040. The upper portion 3040 covers the port 3022 when the secondary cover 3010 is in a closed state. The hinge 3042 is connected to the motor 3032. For example, the hinge 3042 includes a first opening 3044 which may be used to connect the secondary cover 3010 to the drive shaft of the motor 3032. The hinge 3042 may also include a second opening 3046 for attaching an iron element 3028 (e.g., a screw) to which a magnet 3026 on the primary lid 3008 is attached to keep the secondary lid 3010 closed.

[0139] In some embodiments, the secondary cover 3010 includes a plunger 3048 extending from the bottom surface 3050 of the upper portion 3040, such that the plunger is inserted into the port 3022 when the secondary cover 3010 is closed. In addition, a gasket 3024 is fitted around the plunger 3048 to seal the port 3022 and reduce the vaporization of the diluent.

[0140] Figure 39 is an isometric view of the inside of the cell analysis system 100, showing the dispensing system 106 positioning the probe 108 behind the liquid storage module 324. Figure 40 is an isometric view of the inside of the cell analysis system 100, showing the dispensing system 106 and the liquid storage module 324 with the secondary lid 3010 in the open position. The design of the liquid storage module 324, including its slim profile when the primary lid 3008 is in the closed position and the secondary lid 3010 which automatically opens and closes as needed, allows the dispensing system 106 to move the probe 108 freely above the work platform 300 inside the cell analysis system 100, and as a result the probe 108 can access laboratory equipment installed behind the liquid storage module 324 without colliding with the liquid storage module 324. Furthermore, the design of the liquid storage module 324 allows the dispensing system 106 to fully extend probes 108, such as 50 μL or 230 μL pipettes, through port 3022, reaching the bottom of the internal reservoir 3006, and aspirate the diluent, while also allowing the secondary lid 3010 to close and seal port 3022 after the diluent has been aspirated, thereby reducing the vaporization of the diluent during use of the cell analysis system 100.

[0141] The various embodiments described above are provided for illustrative purposes only and should not be construed as limiting in any way. Various modifications can be made to the embodiments described above without departing from the true spirit and scope of this disclosure.

[0142] Embodiments of the present disclosure can be described with reference to the following numbered appendices, which include preferred features developed within the dependent appendices. 1. A fluorescence polarization method for measuring the concentration of antibodies in a sample, wherein the method is: The measurement cycle involves measuring a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, wherein at least one sampling phase of light that fluoresces in the second direction occurs between the sampling phases of light that fluoresces in the first direction. Calculating the first value of the first set in the sampling phase, Calculate the second value of the second set in the sampling phase, The antibody concentration is determined based on a function of the first and second values. Methods that include... 2. The method according to Appendix 1, wherein the first sampling phase of the first set of sampling phases of light that fluoresces in the first direction occurs before the midpoint of the measurement cycle, and the second sampling phase of the first set of sampling phases of light that fluoresces in the first direction occurs after the midpoint of the measurement cycle. 3. The method according to Appendix 1, wherein each sampling phase includes a voltage measurement of fluorescent light emitted from multiple light pulses emitted by a light source. 4. Obtain multiple voltage measurements from each light pulse. The method described in Appendix 3, further including the method described in Appendix 3. 5. The method according to Appendix 4, wherein 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. 6. Calculate the first average value of the first set of voltage measurements, Calculate the second average value of the second set of voltage measurements, The optical pulse difference value is determined for each optical pulse by subtracting the second average value from the first average value. The method described in Appendix 5, further including the method described in Appendix 5. 7. The method according to Appendix 6, further comprising determining the average optical pulse difference with respect to the sampling phase by calculating the average value of the optical pulse difference from the sampling phase. 8. Determine a first polarization value by calculating the average value of the average light pulse difference for a first set of sampling phases of light that is fluorescently emitted in a first direction. The method described in Appendix 7, further including the method described in Appendix 7. 9. Determine the second polarization value by calculating the average value of the average light pulse difference for a second set of sampling phases of light that is fluorescently emitted in the second direction. The method described in Appendix 8, further including the method described in Appendix 8. 10. The method according to Appendix 9, further comprising determining the concentration value by subtracting the second polarization value from the first polarization value and dividing by the sum of the first and second polarization values. 11. The antibody is immunoglobulin G (IgG), as described in Appendix 1. 12. A fluorescence polarization system for measuring the concentration of antibodies in a sample, Light source and A first polarizing filter that polarizes light emitted from a light source in a first direction, A container for holding a sample to be mixed with a fluorescence polarizing assay agent, wherein the container receives light polarized in a first direction, A movable second and third polarizing filter, wherein the second polarizing filter restricts the passage of fluorescent light emitted from inside the container to a first direction, and the third polarizing filter restricts the passage of fluorescent light emitted from inside the container to a second direction, the second being substantially perpendicular to the first direction, and the movable second and third polarizing filters, A detector for measuring light that fluoresces in a first direction and light that fluoresces in a second direction, A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network... The measurement cycle involves measuring a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, wherein at least one sampling phase of light that fluoresces in the second direction occurs between the sampling phases of light that fluoresces in the first direction. Calculating the first value of the first set in the sampling phase, Calculate the second value of the second set in the sampling phase, The antibody concentration is determined based on a function of the first and second values. A processing network that performs this task A system that includes these features. 13. The system as described in Appendix 12, wherein the first sampling phase of the first set of sampling phases of light that fluoresces in the first direction occurs before the midpoint of the measurement cycle, and the second sampling phase of the first set of sampling phases of light that fluoresces in the first direction occurs after the midpoint of the measurement cycle. 14. The system as described in Appendix 13, in which each sampling phase includes a voltage measurement of fluorescent light emitted from multiple light pulses emitted by a light source. 15. When an instruction is executed by the processing network, further, The system described in Appendix 14, which causes a processing circuit network to acquire multiple voltage measurements from each optical pulse. 16. The system as described in Appendix 15, wherein 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. 17. When an instruction is executed by the processing network, it further sends to the processing network: Calculate the first average value of the first set of voltage measurements, Calculate the second average value of the second set of voltage measurements, The optical pulse difference value is determined for each optical pulse by subtracting the second average value from the first average value. The system described in Appendix 16, which enables the following. 18. When an instruction is executed by the processing network, it further sends to the processing network: The system described in Appendix 17, which determines the average optical pulse difference for the sampling phase by calculating the average value of the optical pulse difference from the sampling phase. 19. When an instruction is executed by the processing network, it further sends to the processing network: The system according to Appendix 18, which causes the system to determine a first polarization value by calculating the average value of the average light pulse difference with respect to a first set of sampling phases of light that is fluorescently emitted in a first direction. 20. When an instruction is executed by the processing network, it further sends to the processing network: The system according to Appendix 19, which causes the system to determine a second polarization value by calculating the average value of the average light pulse difference with respect to a second set of sampling phases of light that is fluorescently emitted in a second direction. 21. When an instruction is executed by the processing network, it further sends to the processing network: The system described in Appendix 20 determines the concentration value by subtracting the second polarization value from the first polarization value and dividing by the sum of the first and second polarization values. 22. The antibody is immunoglobulin G (IgG), as described in Appendix 12. 23. A fluorescence polarization method for measuring the concentration of antibodies in a cell sample, the method being: This involves performing a measurement cycle of light emitted from a cell sample mixed with a fluorescence polarizing assay agent, the measurement cycle consisting of the following measurements in this order: (1) Before the midpoint of the measurement cycle, measure the first sampling phase of the fluorescent light emitted in the first direction, (2) Before the midpoint of the measurement cycle, the second sampling phase of the fluorescent light emitted in the second direction is measured, wherein the second direction is perpendicular to the first direction. (3) After the midpoint of the measurement cycle, measure the third sampling phase of the light that is emitted fluorescence in the second direction, (4) After the midpoint of the measurement cycle, measure the fourth sampling phase of the light that is fluorescently emitted in the first direction. This includes, Calculate the first mean value for the first and fourth sampling phases, Calculate the second mean value for the second and third sampling phases, The antibody concentration is determined based on a function of the first and second mean values. Methods that include... 24. The method as described in Appendix 23, wherein each sampling phase includes a voltage measurement of fluorescent light emitted from multiple light pulses emitted by a light source. 25. Obtain multiple voltage measurements from each light pulse. The method described in Appendix 24, further including the method described in Appendix 24. 26. The method according to Appendix 25, wherein 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. 27. Calculate the average value of the first set of voltage measurements, Calculate the average value of the second set of voltage measurements, The optical pulse difference value for each optical pulse is determined by subtracting the average value of the second set of voltage measurements from the average value of the first set of voltage measurements. The method described in Appendix 26, which further includes the method described in Appendix 26. 28. Determine the average voltage value for each sampling phase by calculating the average value of the optical pulse difference between multiple optical pulses in each sampling phase. The method described in Appendix 27, further including the method described in Appendix 27. 29. Determine a first polarization value for a first set of measurements in the sampling phase by calculating the average of the average voltage values ​​of a first set of measurements in the sampling phase. The method described in Appendix 28, further including the method described in Appendix 28. 30. Determine the second polarization value for the second set of measurements in the sampling phase by calculating the average of the average voltage values ​​of the second set of measurements in the sampling phase. The method described in Appendix 29, further including the method described in Appendix 29. 31. Determine the concentration value by subtracting the second voltage value from the first voltage value and dividing by the sum of the first and second voltage values. The method described in Appendix 30, further including the method described in Appendix 30. 32. The antibody is immunoglobulin G (IgG), as described in Appendix 23. 33. A fluorescence polarization system for measuring the concentration of antibody proteins in a cell sample, wherein the system is A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network... This involves performing a measurement cycle of light emitted from a cell sample, the measurement cycle consisting of the following steps: (1) Before the midpoint of the measurement cycle, measure the first sampling phase of the fluorescent light emitted in the first direction, (2) Before the midpoint of the measurement cycle, the second sampling phase of the fluorescent light emitted in the second direction is measured, wherein the second direction is perpendicular to the first direction. (3) After the midpoint of the measurement cycle, measure the third sampling phase of the light that is emitted fluorescence in the second direction, (4) After the midpoint of the measurement cycle, measure the fourth sampling phase of the light that is fluorescently emitted in the first direction. This includes, Calculate the first mean value for the first and fourth sampling phases, Calculate the second mean value for the second and third sampling phases, The antibody concentration is determined based on a function of the first and second mean values. A system equipped with a processing circuit network to perform the following actions. 34. The system as described in Appendix 33, wherein each sampling phase includes a voltage measurement of fluorescent light emitted from multiple light pulses emitted by a light source. 35. When an instruction is executed by the processing network, it further sends to the processing network: Obtaining multiple voltage measurements from each light pulse. The system described in Appendix 34, which enables the following. 36. The system as described in Appendix 35, wherein 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. 37. When an instruction is executed by the processing network, it further sends to the processing network: Calculate the average value of the first set of voltage measurements, Calculate the average value of the second set of voltage measurements, The optical pulse difference value for each optical pulse is determined by subtracting the average value of the second set of voltage measurements from the average value of the first set of voltage measurements. The system described in Appendix 36, which enables the following. 38. When an instruction is executed by the processing network, it further sends to the processing network: The system described in Appendix 37, which determines the voltage value for each sampling phase by calculating the average value of the optical pulse difference between multiple optical pulses in each sampling phase. 39. When an instruction is executed by the processing network, it further sends to the processing network: The system described in Appendix 38, which determines a first polarization value by calculating the average value of the voltage values ​​for each sampling phase of light that is fluorescently emitted in a first direction. 40. When an instruction is executed by the processing network, it is further sent to the processing network. The system described in Appendix 39, which determines a second polarization value by calculating the average value of the voltage values ​​for each sampling phase of light that is fluorescently emitted in a second direction. 41. When an instruction is executed by the processing network, it is further sent to the processing network. The system described in Appendix 40, which determines the antibody concentration by subtracting the second polarization value from the first polarization value and dividing by the sum of the first and second polarization values. 42. The antibody is immunoglobulin G (IgG), as described in Appendix 33. 43. A fluorescence polarization system for measuring the concentration of immunoglobulin G (IgG) in a sample, wherein the system is A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network... The measurement cycle involves measuring a first set of sampling phases of light that fluoresces in a first direction and a second set of sampling phases of light that fluoresces in a second direction, wherein the second direction is substantially perpendicular to the first direction, and at least one sampling phase of light that fluoresces in the second direction occurs between the sampling phases of light that fluoresces in the first direction, and each sampling phase in the first and second sets of sampling phases includes a measurement of the voltage of light from a plurality of light pulses emitted by a light source. Obtaining multiple voltage measurements 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. Calculate the average value of the first set of voltage measurements, Calculate the average value of the second set of voltage measurements, The optical pulse difference value for each optical pulse is determined by subtracting the average value of the second set of voltage measurements from the average value of the first set of voltage measurements, The voltage value for each sampling phase is determined by calculating the average value of the optical pulse difference between multiple optical pulses in each sampling phase, The first polarization value for the first set of sampling phases is determined by calculating the average value of the voltage values ​​in the first set of sampling phases, The second polarization value for the second set of sampling phases is determined by calculating the average value of the voltage values ​​for the second set of sampling phases, The IgG concentration value is determined by subtracting the second polarization value from the first polarization value and dividing by the sum of the first and second polarization values. A system equipped with a processing circuit network to perform the following actions. 44. A cell analysis system for measuring the concentration of antibodies in a sample, wherein the system is Light source and An excitation polarizer that polarizes light emitted from a light source in a first direction, A container configured to receive light polarized in a first direction, A polarizer holder for holding a first detection polarizer and a second detection polarizer, wherein the first detection polarizer restricts the passage of fluorescent light emitted from inside the container in a first direction, and the second detection polarizer restricts the passage of fluorescent light emitted from inside the container in a second direction, the second direction being perpendicular to the first direction, and the polarizer holder. A detector for measuring light that fluoresces in a first direction and light that fluoresces in a second direction, A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network... A processing circuit network is used to rotate the polarizer holder around the axis of rotation in alternating directions, causing the first and second detection polarizers to be alternately positioned within the optical path of the fluorescent light emitted from inside the container. A system that includes these features. 45. When an instruction is executed by the processing network, it is further sent to the processing network. The polarizer holder is rotated to position the first detection polarizer within the optical path of the fluorescent light emitted from inside the container, The polarizer holder is rotated to position the second detection polarizer within the optical path of the fluorescent light emitted from inside the container. The system described in Appendix 44, which enables the following. 46. ​​Further comprising an optical plate having grooves that terminate within a first stop and a second stop, The polarizer holder is part of the system described in Appendix 44, which includes a pin that displaces inward into a groove in the optical plate as the polarizer holder rotates about an axis of rotation. 47. The optical plate further includes a first magnet positioned next to a first stop in the groove and a second magnet positioned next to a second stop. The pin is made of a ferromagnetic material that, when positioned next to the first stop at the end of the first rotational progression, is attracted by the first magnet, and the pin, when positioned next to the second stop at the end of the second rotational progression, is attracted to the second magnet. The system described in Appendix 46. 48. The groove is curved along the axis of rotation, as described in Appendix 46. 49. The polarizer holder comprising a first side surface and a second side surface perpendicular to the first side surface, wherein the first side surface comprises a first recess for holding a first detection polarizer, and the second side surface comprises a second recess for holding a second detection polarizer, as described in Appendix 44. 50. The polarizer holder further includes a first biasing element attached to a first side surface for fixing a first detection polarizer within a first recess, and a second biasing element attached to a second side surface for fixing a second detection polarizer within a second recess, as described in Appendix 49. 51. The system as described in Appendix 50, wherein the first detection polarizer, when installed inside the first recess, protrudes beyond the first side surface, the second detection polarizer, when installed inside the second recess, protrudes beyond the second side surface, the first biasing element flexes around the first detection polarizer, fixing the first detection polarizer in place, and the second biasing element flexes around the second detection polarizer, fixing the second detection polarizer in place. 52. The system according to Appendix 51, wherein the first biasing element and the second biasing element include cutouts for increasing the flexure around the first and second detection polarizers. 53. The polarizer holder further includes a curved surface connecting a first side surface and a second side surface, the curved surface having an opening, the opening aligning with openings in the first recess and the second recess, allowing light emitted fluorescently from within the container to pass through the polarizer holder and reach the detector, as described in Appendix 49. 54. The system as described in Appendix 53, further comprising an optical cover that at least partially houses the detector and the polarizer holder, the optical cover including a curved wall, the curved wall aligning with the curved surface of the polarizer holder and reducing the space between the polarizer holder and the curved wall. 55. Work platform and, A liquid storage module mounted on a work platform, wherein the liquid storage module is A base having an internal volume for holding the internal reservoir, A primary cover connected to the base, the primary cover includes a port providing access to an internal reservoir, A secondary cover that is attached to a primary cover, the secondary cover is configured to seal the ports on the primary cover, A second motor is attached to the secondary cover, A liquid storage module, A dispensing system configured to move a probe along three mutually perpendicular axes while positioned above the work platform. Furthermore, When an instruction is executed by the processing network, it is further sent to the processing network. The secondary lid is opened, and the second motor is operated to allow the dispensing system probe to be inserted through the port on the primary lid, reach the bottom of the internal reservoir, and aspirate the liquid contained therein. Once the probe is removed from the port, the secondary cover is closed and the second motor is operated to seal the port on the primary cover. The system described in Appendix 44, which enables the following. 56. A polarizer holder for a cell analysis system, wherein the polarizer holder is The first side surface and 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 a first side surface, which restricts the passage of fluorescently emitted light in a first direction, A second detection polarizer coupled to a second side surface, the second detection polarizer comprises a second detection polarizer that restricts the passage of fluorescent light in a second direction, The polarizer holder rotates about a rotation axis and is configured to cause alternating placement of the first and second detection polarizers within the optical path of light. Polarizer holder. 57. A first recessed portion within the first side surface, wherein the first detection polarizer is mounted within the first recessed portion, and the first recessed portion and A second recessed portion within the second side surface, the second detection polarizer is mounted within the second recessed portion and A polarizer holder further equipped with the features described in Appendix 56. 58. A first biasing element attached to the first side surface for fixing the first detection polarizer within the first recessed portion, A second biasing element is attached to the second side surface to fix the second detection polarizer within the second recessed portion. A polarizer holder further equipped with the features described in Appendix 57. 59. A polarizer holder as described in Appendix 58, wherein the first detection polarizer, when installed inside the first recess, protrudes beyond the first side surface, the second detection polarizer, when installed inside the second recess, protrudes beyond the second side surface, the first biasing element flexes around the first detection polarizer, fixing the first detection polarizer in place, and the second biasing element flexes around the second detection polarizer, fixing the second detection polarizer in place. 60. A polarizer holder as described in Appendix 59, wherein the first and second retaining springs include cutouts for increasing the flexure around the first and second detection polarizers. 61. A fluorescence polarization method for measuring the concentration of antibodies in a sample, The polarizer holder is positioned in a first position such that the first detection polarizer is placed within the optical path of light that fluoresces from within a container holding a sample of antibody mixed with a fluorescence polarizing assay agent. The method involves detecting light that fluoresces from inside the container after light has passed through a first detection polarizer, wherein the first detection polarizer restricts the passage of the fluorescent light from inside the container in a first direction. The polarizer holder is rotated around its axis of rotation so that it is positioned in a second location, such that the second detection polarizer is placed in the optical path of the light emitted fluorescently from inside the container, and the second detection polarizer restricts the passage of the light emitted fluorescently from inside the container in a second direction. The process involves detecting the light that is emitted fluorescently from inside the container after the light has passed through a second detection polarizer. Methods that include... 62. The method according to Appendix 61, further comprising rotating the polarizer holder about its axis of rotation so as to return the polarizer holder from a second position to a first position. 63. The polarizer holder is rotated 90 degrees between the first position and the second position, as described in Appendix 61. 64. The method according to Appendix 61, further comprising determining the concentration of an antibody based on detecting light passing through a first detection polarizer and a second detection polarizer. 65. A module for a cell analysis system, the module is A base having an internal volume that holds the internal reservoir, A primary cover connected to the base, the primary cover includes a port providing access to an internal reservoir, A secondary cover that is attached to a primary cover, the secondary cover is configured to seal the ports on the primary cover, A motor that is attached to the secondary cover, A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network... The motor is operated to open the secondary cover, allowing the probe to be inserted through the port on the primary cover and reach the bottom of the internal reservoir. Once the probe is removed, the motor is operated to close the secondary cover and seal the port on the primary cover. A processing network that performs this task A module equipped with [the following features]. 66. The module described in Appendix 65, further comprising a gasket that seals around the internal reservoir when the primary cover is closed. 67. The module described in Appendix 65, further comprising a gasket that seals the port on the primary cover when the secondary cover is closed. 68. The secondary cover includes an upper portion that covers the ports on the primary cover when the secondary cover is closed, and a hinge extending from the upper portion, the hinge being connected to a motor, as described in Appendix 65. 69. The module according to Appendix 65, wherein the secondary cover includes at least one of a magnet attracted to a corresponding element on the primary cover and an iron element in order to maintain the secondary cover in a closed state. 70. The module as described in Appendix 65, wherein the secondary cover includes a plunger, the plunger being configured to be inserted at least partially into a port on the primary cover.

Claims

1. A system for measuring the concentration of antibodies in a sample, wherein the system is Light source and An excitation polarizer that polarizes the light emitted from the light source in a first direction, A container configured to receive the light polarized in the first direction, A polarizer holder for holding a first detection polarizer and a second detection polarizer, wherein the first detection polarizer restricts the passage of light emitted fluorescently from within the container in a first direction, and the second detection polarizer restricts the passage of the light emitted fluorescently from within the container in a second direction, the second direction being perpendicular to the first direction, and the polarizer holder, A detector for measuring the light that fluoresces in the first direction and the light that fluoresces in the second direction, A processing network, wherein the processing network has a memory for storing instructions, and when an instruction is executed by the processing network, the processing network receives A processing circuit network is provided to rotate the polarizer holder around the axis of rotation in alternating directions, causing the first and second detection polarizers to be alternately positioned within the optical path of the light emitted fluorescently from inside the container. A system that includes these features.

2. When the aforementioned instruction is executed by the processing network, the processing network further: Rotating the polarizer holder to position the first detection polarizer within the optical path of the light emitted fluorescently from inside the container, Rotating the polarizer holder to position the second detection polarizer within the optical path of the light that is fluorescently emitted from inside the container. The system according to claim 1, which causes the following to be performed.

3. The optical plate further comprises a groove that terminates within a first stop and a second stop, The system according to claim 1, wherein the polarizer holder includes a pin that is displaced inward from the groove of the optical plate as the polarizer holder rotates about the axis of rotation.

4. The optical plate further includes a first magnet positioned next to the first stop portion of the groove, and a second magnet positioned next to the second stop portion. The pin comprises a ferromagnetic material which, when positioned next to the first stop at the end of the first rotational progression, is attracted by the first magnet, and the pin which, when positioned next to the second stop at the end of the second rotational progression, is attracted to the second magnet. The system according to claim 3.

5. The system according to claim 3, wherein the groove is curved along the axis of rotation.

6. The polarizer holder comprises a first side surface and a second side surface perpendicular to the first side surface, wherein the first side surface comprises a first recess for holding the first detection polarizer, and the second side surface comprises a second recess for holding the second detection polarizer, according to claim 1.

7. The system according to claim 6, wherein the polarizer holder further includes a first biasing element attached to the first side surface for fixing the first detection polarizer within the first recess, and a second biasing element attached to the second side surface for fixing the second detection polarizer within the second recess.

8. The system according to claim 7, wherein the first detection polarizer, when installed inside the first recessed portion, protrudes beyond the first side surface; the second detection polarizer, when installed inside the second recessed portion, protrudes beyond the second side surface; the first biasing element flexes around the first detection polarizer, fixing the first detection polarizer in place; and the second biasing element flexes around the second detection polarizer, fixing the second detection polarizer in place.

9. The system according to claim 8, wherein the first biasing element and the second biasing element include cutouts for increasing the flexure around the first and second detection polarizers.

10. The polarizer holder further includes a curved surface connecting the first side surface and the second side surface, the curved surface having an opening, the opening aligning with openings in the first recess and the second recess, allowing the light emitted fluorescently from within the container to pass through the polarizer holder and reach the detector, according to claim 6.

11. The system according to claim 10, further comprising an optical cover that at least partially houses the detector and the polarizer holder, the optical cover including a curved wall, the curved wall aligning with the curved surface of the polarizer holder and reducing the space between the polarizer holder and the curved wall.

12. Work platform and A liquid storage module mounted on the aforementioned work platform, wherein the liquid storage module is A base having an internal volume for holding the internal reservoir, A primary cover connected to the base, the primary cover includes a port providing access to the internal reservoir, A secondary cover attached to the primary cover, wherein the secondary cover is configured to seal the port on the primary cover, A second motor attached to the secondary cover and A liquid storage module, A dispensing system configured to move a probe along three mutually perpendicular axes above the aforementioned work platform. Furthermore, When the aforementioned instruction is executed by the processing network, the processing network further: The second motor is operated to open the secondary lid, allowing the probe of the dispensing system to be inserted through the port on the primary lid, reach the bottom of the internal reservoir, and aspirate the liquid contained therein. When the probe is removed from the port, the second motor is operated to close the secondary cover and seal the port on the primary cover. The system according to claim 1, which causes the following to be performed.

13. A polarizer holder for a cell analysis system, wherein the polarizer holder is The first side surface and 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 restricts the passage of fluorescent light in a first direction, A second detection polarizer coupled to the second side surface, wherein the second detection polarizer restricts the passage of fluorescently emitted light in a second direction. Equipped with, The polarizer holder is configured to rotate about a rotation axis, causing the first detection polarizer and the second detection polarizer to be positioned alternately within the optical path of the light.

14. The first recessed portion within the first side surface, the first detection polarizer is mounted within the first recessed portion, and The second recessed portion within the second side surface, the second detection polarizer is mounted within the second recessed portion, and The polarizer holder according to claim 13, further comprising:

15. A first biasing element is attached to the first side surface to fix the first detection polarizer within the first recessed portion, A second biasing element is attached to the second side surface to fix the second detection polarizer within the second recessed portion. The polarizer holder according to claim 14, further comprising:

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