Information processing apparatus, biological sample analysis method, biological sample detection apparatus, and biological sample detection system

Abstract

To provide a technology for supporting the selection of a binding molecule and a labeled molecule to be used to detect and / or analyze a target molecule.SOLUTION: An information processing device has a processing part for calculating reactivities of a target molecule and a plurality of different binding molecules in using the plurality of different binding molecules labeled with different labeled molecules on the basis of signals derived from a sample including a biological specimen is provided. The signal includes a first signal group acquired when the plurality of different binding molecules labeled with the same kind of a labeled molecule are reacted with the target molecule, and a second signal group acquired when the same kind of binding molecules labeled with different labeled molecules are reacted with the target molecule.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present technology relates to an information processing apparatus. More specifically, it relates to an information processing apparatus used for detecting a biological sample, a biological sample analysis method, a biological sample detection apparatus, and a biological sample detection system.

Background Art

[0002] For analyzing various molecules, various analyses using labels are being performed. For example, molecules such as antigen proteins are detected and / or analyzed using antibodies labeled with multiple fluorescent dyes by a flow cytometer or a microscope. In addition to antigen-antibody reactions, detection and analysis of molecules by nucleic acid hybridization using fluorescently labeled nucleic acid probes and detection and analysis of enzyme molecules using fluorescently labeled substrates are widely performed. In these detections and / or analyses, various fluorescent dyes are used. Fluorescent dyes each have unique properties, such as unique fluorescence spectra and fluorescence intensities.

[0003] For example, Patent Document 1 describes an invention related to a technique for analyzing the type of fluorescence emitted from microparticles (paragraph 0001). The following Patent Document 1 describes "a data display method for displaying a fluorescence spectrum obtained by integrating or averaging detection data obtained by simultaneously detecting fluorescence emitted from microparticles flowing through a flow path in a plurality of wavelength regions for a plurality of microparticles." (Claim 1).

[0004] Also, Patent Document 2 describes a method for presenting combinations of antibodies, nucleic acid probes, etc. that bind to antigens, nucleic acids, etc. and fluorescent dyes.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0006] When a binding molecule such as an antibody or a nucleic acid probe is reacted with a target molecule such as an antigen or a nucleic acid, the reactivity varies depending on the types of the target molecule and the binding molecule. Also, the reactivity of the binding molecule with respect to the target molecule varies depending on the type of a labeling molecule such as fluorescence for labeling the binding molecule. For this reason, the detection accuracy is also affected by the types of the binding molecule and the labeling molecule used for detecting the target molecule.

[0007] In order to detect and / or analyze a target molecule, it is necessary to select a binding molecule and a labeling molecule. Usually, the selection of the binding molecule and the labeling molecule is performed by the user himself / herself who performs the detection and / or analysis, but this is a laborious task even if the user is experienced. Also, it cannot be denied that the selected binding molecule and labeling molecule may not be suitable for the detection and / or analysis of the target molecule.

[0008] In Patent Document 2, a method of performing evaluation based on measured autofluorescence and fluorescence single staining and presenting a combination of fluorescent dyes has been proposed. However, in this case, the signal of the target fluorescently labeled antibody needs to be actually measured using the target fluorescently labeled antibody itself, and based on the signal data of the antibody labeled with another fluorescent dye, the signal data of the antibody labeled with another fluorescent dye cannot be accurately calculated. That is, Patent Document 2 does not describe a calculation method for unmeasured fluorescently labeled antibodies.

[0009] Therefore, the main object of the present technology is to provide a technology for assisting the selection of a binding molecule and a labeling molecule used for detecting and / or analyzing a target molecule.

Means for Solving the Problems

[0010] In this technology, first, based on signals from a sample containing a biological sample, a processing unit calculates the reactivity of a target molecule with each of a plurality of different binding molecules labeled with different labeling molecules. It has The signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, and provides an information processing apparatus including the above. In the information processing apparatus according to this technology, as an index of the reactivity, the number of binding molecules and / or the fluorescence intensity labeled on the number of binding molecules can be calculated. In this case, the fluorescence intensity can be calculated from one or more numerical values selected from excitation efficiency, quantum yield, absorption efficiency, and photolabeling rate (F / P value). In this technology, the signal can include at least one of a signal, a specific signal / background, and a specific signal / nonspecific signal. In the processing unit of the information processing apparatus according to this technology, based on a third signal obtained when a negative control is used for a target molecule, the background in each detection channel can also be calculated. In the processing unit of the information processing apparatus according to this technology, the leakage of autofluorescence signals and / or the leakage of signals derived from other labeling molecules can be calculated. In the processing unit of the information processing apparatus according to this technology, the reactivity with a target molecule can also be calculated for a combination of a labeling molecule and a binding molecule that has not actually been measured. In the processing unit of the information processing apparatus according to this technology, a combination of a labeling molecule and a binding molecule for which the specific signal / background is equal to or greater than a threshold value can also be selected. In the processing unit of the information processing apparatus according to the present technology, it is also possible to select a combination of a labeled molecule and a binding molecule that maximizes the sum of the specific signal / background. In the processing unit of the information processing apparatus according to the present technology, it is also possible to select a combination of a labeled molecule and a binding molecule that maximizes the sum of the difference between the signal of the fluorescence intensity and the background of the fluorescence intensity. In the processing unit of the information processing apparatus according to the present technology, it is also possible to select a combination of a labeled molecule and a binding molecule based on the magnitude of the signal of the fluorescence intensity. In the processing unit of the information processing apparatus according to the present technology, it is also possible to select a combination of a labeled molecule and a binding molecule such that the binding molecules with small values in the first signal group are assigned in order, and the labels with high fluorescence intensity signals are assigned in order. In the processing unit of the information processing apparatus according to the present technology, it is also possible to select a combination of a labeled molecule and a binding molecule such that the binding molecules with large values in the first signal group are assigned in order, and the labels with short detection wavelengths are assigned in order. The information processing apparatus according to the present technology may further include a presentation unit that presents support information on a combination of a binding molecule and a labeled molecule to the user based on the calculated reactivity. Further, the information processing apparatus according to the present technology may further include an evaluation unit that estimates the significance of a binding molecule and / or a labeled molecule with respect to a target molecule based on image information. In the present technology, the first signal group and the second signal group can be detection amounts normalized using at least one selected from excitation power density, exposure time, and detection device sensitivity.

[0011] In the present technology, next, a signal acquisition step of acquiring a signal derived from a sample containing a biological sample, a processing step of calculating the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeled molecules based on the signal, an output step of outputting the reactivity, and the signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, and A biological sample analysis method including the above is provided.

[0012] In this technology, further, a signal acquisition unit that acquires a signal derived from a sample including a biological sample, A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal, An output unit that outputs the reactivity, A detection unit that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity, and has The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, and A biological sample detection device including the above is provided. The biological sample detection device according to this technology can further include a labeling unit that labels a target molecule using a binding molecule labeled with a labeling molecule selected based on the output reactivity. The biological sample detection device according to this technology can further include an analysis unit that analyzes the sample based on the signal detected by the detection unit.

[0013] In this technology, furthermore, a signal acquisition unit that acquires a signal derived from a sample including a biological sample, A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal; An output unit that outputs the reactivity, and the signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, An information processing apparatus including; A detection apparatus that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity, and A biological sample detection system including is provided.

Brief Description of Drawings

[0014]

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Mode for Carrying Out the Invention

[0015] Hereinafter, a preferred mode for carrying out the present technology will be described with reference to the drawings. The embodiments described below show an example of a typical embodiment of the present technology, and the scope of the present technology is not construed narrowly thereby. The description will be made in the following order. 1. Information processing apparatus 1 (1) Target molecule (2) Binding molecule (3) Labeled molecule (4) Signal acquisition unit 11 (5) Processing unit 12 (a) First signal group (b) Second signal group (c) Reactivity calculation: Processing unit 12 (d) Third signal (e) Selection of combination of labeled molecule and binding molecule: Processing unit 12 (6) Evaluation unit 13 (7) Output unit 14 (8) Presentation unit 15 (9) Memory unit 16 (10) Display unit 17 (11) User interface 18 2. Information processing system 2 3. Biological sample detection device 3, biological sample detection system 4 (1) Detection unit 31, detection device 41 (2) Labeling unit 32, labeling device 42 (3) Analysis unit 33, analysis device 43 4. Computer program 5. Biological sample analysis method 6. Application examples [Microscope system 5000] [Biological sample analyzer 6100]

[0016] <1. Information processing device 1> Figure 1 is a block diagram showing a configuration example of the biological sample detection system 4 according to the present technology. The information processing device 1 according to the present technology is an information processing device that can be used in the biological sample detection system 4 according to the present technology described later. The information processing device 1 according to the present technology includes at least a processing unit 12. Further, if necessary, a signal acquisition unit 11, an evaluation unit 13, an output unit 14, a presentation unit 15, a memory unit 16, a display unit 17, a user interface 18, etc. can also be provided. Hereinafter, each unit etc. will be described in detail.

[0017] (1) Target molecule In this technology, a target molecule is a molecule that can be detected and / or analyzed by binding to a binding molecule labeled with a labeling molecule described later, and can be appropriately selected by those skilled in the art. The target molecule is, for example, a molecule that can be detected and / or analyzed by binding to a binding molecule labeled with a labeling molecule in analyses such as flow cytometry, microscopy, Western blot, various arrays, and ELISA. That is, this technology is used to assist in the selection of labeling molecules and binding molecules used in these analyses and can be used for it.

[0018] More specifically, the target molecule is, for example, a molecule that can exist in a living body including body fluids such as blood and urine and tissues, and examples include biomolecules, drug molecules, harmful molecules, etc. Examples of biomolecules include nucleic acids, proteins, saccharides, lipids, vitamins, etc. Examples of nucleic acids include DNA and RNA. Examples of proteins include, for example, antigen proteins, enzyme proteins, structural proteins, adhesion proteins, etc. Furthermore, the target molecule includes those that correlate with changes in diseases and responses to treatments as biomarkers and serve as indicators.

[0019] (2) Binding molecule In this technology, a binding molecule is a molecule that enables the detection and / or analysis of a target molecule by binding to the aforementioned target molecule, and can be appropriately selected by those skilled in the art. The binding molecule is, for example, a molecule that can detect and / or analyze a target molecule by binding to the target molecule in the various analyses described above.

[0020] More specifically, the binding molecule is a molecule that specifically binds to a target molecule, and examples thereof include biomolecules, drug molecules, high molecular weight compounds, low molecular weight compounds, etc. For example, nucleic acids, artificial nucleic acids, proteins, peptides, saccharides, lipids, and vitamins can be mentioned. In addition, DNA and RNA, PNA, LNA, etc. can be mentioned. Further, as examples of antibody-like molecules, antibody cell surface markers, enzyme proteins, structural proteins, adhesion proteins, etc. can be mentioned.

[0021] A method for analyzing a target molecule using an antibody to which a fluorescent dye is bound as a labeling molecule described later as a binding molecule is called fluorescence immunostaining. Fluorescence immunostaining includes, for example, immunocytochemistry (ICC), immunohistochemistry (IHC), etc. ICC is a method for staining cells separated from a tissue or cultured cells. IHC is a method for staining target molecules in thin sections of a tissue.

[0022] In addition, the fluorescence immunostaining includes a direct fluorescence immunostaining method and an indirect fluorescence immunostaining method. The direct fluorescence immunostaining method is a method in which an antibody bound to a fluorescent dye directly binds to a target molecule, and the target molecule is analyzed by detecting the fluorescent dye. In the indirect fluorescence immunostaining method, an antibody bound to a fluorescent dye (also called a secondary antibody) binds to an antibody (also called a primary antibody) that specifically binds to a target molecule, and this further binds to the target molecule. That is, the indirect fluorescence immunostaining method is a method in which an antibody bound to a fluorescent dye (also called a secondary antibody) binds to a target molecule via a primary antibody, and the target molecule is analyzed by detecting the fluorescent dye.

[0023] This technology can be suitably used for the selection of binding molecules and labeling molecules used in fluorescence immunostaining.

[0024] (3) Labeling molecule In this technology, the labeling molecule is a molecule that labels the aforementioned binding molecule. When this binding molecule binds to the target molecule, it enables the detection and / or analysis of the target molecule. Examples of the labeling molecule include molecules that can be used as labeling molecules in the various analyses described above.

[0025] More specifically, examples of the labeling molecule include dyes. Examples of the dyes include various fluorescent dyes having fluorescence wavelengths in the visible light region, such as fluorescent dyes of the AlexaFluor (registered trademark) series, fluorescent dyes of the DyLight (registered trademark) series, and fluorescent dyes of the BD Horizon Brilliant (trademark) series, Atto, FITC, Cy3, Cy5, Cy5.5, Cy7, Rhodamine, PE (phycoerythrin), APC (Allophycocyanin), PerCP, etc., but are not limited thereto.

[0026] In addition, in this technology, the labeling molecule may be one that is expressed as a part of the target molecule or the binding molecule, such as a fluorescent protein contained in a fluorescent fusion protein. Examples of the fluorescent protein include GFP, BFP, CFP, EGFP, EYFP, and PA-GFP.

[0027] (4) Signal acquisition unit 11 In the signal acquisition unit 11, signals derived from a sample containing a biological sample are acquired. For example, signals detected by a detection device 41 such as a flow cytometer, a microscope, or various photodetectors are acquired by the signal acquisition unit 11.

[0028] In the signal acquisition unit 11, not only signals detected by various detection devices 41 but also signal data in a database stored in a storage unit 16 described later can be acquired. For example, past detection data, data detected by other detection devices and accumulated in the database, etc. can also be acquired by the signal acquisition unit 11.

[0029] (5) Processing Unit 12 Based on the signals acquired by the signal acquisition unit 11, the processing unit 12 calculates the reactivity of each of the target molecule with a plurality of different binding molecules labeled with different labeling molecules.

[0030] In this technology, the signal may be the fluorescence signal itself, specific signal / background, specific signal / nonspecific signal, etc. Also, the fluorescence signal may be in pixel units such as signal / pixel, or in cell units such as average fluorescence signal / cell, total fluorescence signal / cell.

[0031] The signal serving as the basis for the calculation performed by the processing unit 12 includes a first signal group and a second signal group. It may also include a third signal. Hereinafter, each signal group and the method for calculating the reactivity performed by the processing unit 12 will be described in detail.

[0032] (a) First Signal Group The first signal group is a group of signals obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule. For example, a plurality of binding molecules C1 - C5 (F1 - C1, F1 - C2, F1 - C3, F1 - C4, F1 - C5) labeled with a labeling molecule F1 are each reacted with a target molecule to obtain respective signals.

[0033] By acquiring the first signal group, the reactivity of the binding molecules C1 - C5 with respect to the target molecule can be evaluated. That is, the reactivity between different binding molecules C1 - C5 with respect to the target molecule can be compared, and the reactivity can be calculated. The index representing the reactivity is not particularly limited as long as the reactivity can be evaluated. For example, fluorescence intensity, number of binding molecules, absorbance, light scattering, phosphorescence, fluorescence lifetime, mass, etc. can be mentioned.

[0034] The method for calculating the number of binding molecules is not particularly limited and can be calculated using general calculation methods. For example, it can be calculated using the antibody number conversion method described in WO2020 / 022038.

[0035] An example of the method for calculating the number of binding molecules will be described with reference to FIG. 2. In step S1000, the user determines the labeling molecule, binding molecule, and target molecule to be used for the analysis. In step S1004, the user creates a stained specimen by staining the target molecule using the labeling molecule and the binding molecule.

[0036] In step S1008, the signal acquisition unit 11 of the information processing apparatus 1 acquires imaging image information by imaging the stained specimen. In step S1012, the signal acquisition unit 11 acquires reagent information such as the fading coefficient, absorption cross-section area, quantum yield, and fluorescence labeling rate from the database of the storage unit 16 based on the reagent identification information attached to the labeling molecule and the binding molecule used for the generation of the stained specimen. In addition, the signal acquisition unit 11 acquires the separately measured excitation power density.

[0037] In step S1016, the processing unit 12 corrects the luminance of each pixel in the imaging image information using the fading coefficient, absorption cross-section area, and excitation power density (fading correction processing is performed). In step S1020, the processing unit 12 converts the luminance of each corrected pixel into the number of photons. In step S1024, the processing unit 12 converts the number of photons into the number of labeling molecules or the number of binding molecules bound to the labeling molecules.

[0038] In step S1028, the processing unit 12 generates image information reflecting the number of labeling molecules or the number of binding molecules bound to the labeling molecules. In step S1032, the display unit 17 displays the image information on the display, and a series of processes ends.

[0039] Also, the method for calculating the fluorescence intensity is not particularly limited and can be calculated using general calculation methods. For example, it can be calculated using one or more numerical values selected from the excitation efficiency, quantum yield, absorption efficiency, and optical labeling rate (F / P value).

[0040] (b) Second signal group The second signal group is the signal obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule. For example, the same type of binding molecules C1 labeled with different labeling molecules F1 to F5 (F1-C1, F2-C1, F3-C1, F4-C1, F5-C1) are each reacted with the target molecule to obtain each signal.

[0041] By obtaining the second signal group, it is possible to evaluate the difference in the reactivity of the binding molecules to the target molecule due to the difference in the types of labeling molecules that label the binding molecules. That is, the reactivity between the binding molecules labeled with different labeling molecules F1 to F5 and the target molecule can be compared and evaluated. As an index representing the reactivity, similar to the first signal group, it is not particularly limited as long as the reactivity can be evaluated. For example, the number of binding molecules, fluorescence intensity, etc. can be mentioned.

[0042] Since the method for calculating the number of binding molecules and the method for calculating the fluorescence intensity are the same as those described for the first signal group, the description is omitted here.

[0043] (c) Calculation of reactivity: Processing unit 12 In this technology, based on the first signal group and the second signal group described above, the reactivity between the target molecule and each of a plurality of different binding molecules C2 to C5 labeled with different labeling molecules F2 to F5 is calculated.

[0044] Specifically, since the reactivity of the binding molecules C1 to C5 labeled with the labeling molecule F1 to the target molecule can be evaluated by the first signal group, for example, an index (such as the number of binding molecules, fluorescence intensity, etc.) representing the reactivity of the binding molecules C1 to C5 to the target molecule can be displayed in the column of the labeling molecule F1 in Table 1 below (see the vertical line in Table 1).

[0045] In addition, since the second signal group can evaluate the difference in the reactivity of the binding molecule C1 with respect to the target molecule due to the difference in the types of the labeling molecules F1 to F5 that label the binding molecule C1, for example, in the row of the binding molecule C1 in Table 1 below, an index (for example, the number of binding molecules, fluorescence intensity, etc.) representing the reactivity of the binding molecule C1 labeled with the labeling molecules F1 to F5 with respect to the target molecule can be displayed (see the horizontal line in Table 1).

[0046] Then, based on the first signal group and the second signal group, the reactivity of each of the target molecule with a plurality of different binding molecules C2 to C5 labeled with different labeling molecules F2 to F5 is calculated. Specifically, for example, from the reactivity of the binding molecule C1 labeled with the labeling molecule F1 with respect to the target molecule and the reactivity of the binding molecule C2 labeled with the labeling molecule F1 with respect to the target molecule, an estimate of the coefficient of the difference in the reactivity between the binding molecules C1 and C2 with respect to the target molecule is made. On the other hand, from the reactivity of the binding molecule C1 labeled with the labeling molecule F1 with respect to the target molecule and the reactivity of the binding molecule C1 labeled with the labeling molecule F2 with respect to the target molecule, an estimate of the coefficient of the difference in the reactivity between the target molecule and the binding molecule C1 when labeled with the labeling molecule F1 and when labeled with the labeling molecule F2 is made. Taking these coefficients into account, the reactivity of the binding molecule C2 labeled with the labeling molecule F2 with respect to the target molecule can be calculated.

[0047] By calculating the reactivity in the same way for the blank portions in Table 1 below, a matrix can be created for the reactivity of each of a plurality of different binding molecules C1 to C5 labeled with different labeling molecules F1 to F5.

[0048]

Table 1

[0049] The reactivity index of the first signal group and the reactivity index of the second signal group may be different. For example, when the number of bound molecules is used for the first signal group and the fluorescence intensity is used for the second signal group, a specific example of a method for calculating the reactivity index of the target molecule and the binding molecule in a combination of labeled molecules and binding molecules that are not actually measured is shown below.

[0050] As the first signal group, the number of bound molecules calculated from imaging images and the like obtained when a plurality of different binding molecules C1 to C3 labeled with the same type of labeled molecule F1 are each reacted with the target molecule are shown in Table 2 below.

[0051]

Table 2

[0052] As the second signal group, the fluorescence intensities calculated from imaging images and the like obtained when the same type of binding molecule C1 labeled with different labeled molecules F1 to F4 are each reacted with the target molecule are shown in Table 3 below. At this time, as will be described later, the signal / background ratio of the fluorescence intensity can also be used.

[0053]

Table 3

[0054] At this time, for example, when the fluorescence intensity is used as the reactivity index, the fluorescence intensities of the labeled molecule F4 and the binding molecule C3 are not actually measured, but can be calculated using the following formula. Reactivity index (fluorescence intensity) of F4-C3 = (Number of bound molecules of F1-C3 / Number of bound molecules of F1-C1) × Fluorescence intensity of F4-C1

[0055] In the same way, by calculating the reactivity index for the blank part of Table 3 above, a matrix can be created for the respective fluorescence intensities when using a plurality of different binding molecules C1 to C3 labeled with different labeled molecules F1 to F4.

[0056] By referring to the matrix created in this way, when selecting binding molecules and labeling molecules to be used in detecting and / or analyzing a target molecule, it becomes possible to select a more optimal combination of binding molecules and labeling molecules. As a result, the accuracy of detecting and / or analyzing the target molecule can be improved.

[0057] As described above, in the present technology, since the reactivity of each of the types of labeling molecules F2 to F5 and the types of binding molecules C2 to C5 that are not actually measured can be calculated based on the first signal group and the second signal group, it is not necessary to actually measure all combinations of binding molecules and labeling molecules. That is, in the present technology, the reactivity of each of the types of labeling molecules and the types of binding molecules can be calculated from a small amount of actually measured data. In other words, the processing unit 12 can calculate the reactivity with the target molecule for combinations of labeling molecules and binding molecules that are not actually measured.

[0058] In addition, by using the present technology, the labor of the user in selecting binding molecules and labeling molecules to be used in detecting and / or analyzing a target molecule can be saved, and it is possible to prevent variations in the accuracy of detecting and / or analyzing the target molecule depending on the user's experience.

[0059] (d) The third signal The third signal is a signal obtained when using a negative control. Examples of signals obtained when using a negative control include a signal obtained when reacting an unlabeled binding molecule, a signal obtained from an unstained specimen without using a binding molecule, a signal obtained when using an isotype control antibody, and the like.

[0060] Based on the third signal, the background in each detection channel can be calculated. Specifically, as the background in each detection channel, it is possible to calculate, for example, the leakage of the autofluorescence signal, the leakage of signals from other labeled molecules such as other fluorescent dye signals, and the like.

[0061] In calculating the background in each detection channel, it is preferable to take into account the leakage from labeled molecules other than the self-channel. The leakage from labeled molecules other than the self-channel can be, for example, the sum of the leakage in each labeled molecule calculated from the second signal group (the signal group obtained when the same type of binding molecules labeled with different labeled molecules are each reacted). Also, for example, the leakage into the self-channel when using binding molecules labeled with all labeled molecules other than the self-channel may be calculated.

[0062] For the first signal group, the second signal group, and the third signal, the detected amounts normalized using at least one selected from the excitation power density, exposure time, and detection device sensitivity can be used. For example, when using a common device, device-to-device correction is possible by management at the time of shipment, and even for errors due to aging deterioration or usage environment, device-to-device or in-device calibration can be performed in daily maintenance. However, even when using different devices or when device-to-device or in-device calibration cannot be performed at the time of shipment or in daily maintenance, it is possible to align the measurement conditions by performing normalization using at least one selected from the excitation power density, exposure time, and detection device sensitivity.

[0063] Also, as will be described later, by accumulating data such as the first signal group, the second signal group, the third signal, and the background and creating a database, the accuracy of device-to-device correction can be further improved according to the tendency (correction coefficient) of device-to-device errors.

[0064] In this technology, as described above, fluorescence intensity can be used as an index representing reactivity, but it is preferable to use the signal / background ratio of fluorescence intensity as an index. By using the signal / background ratio of fluorescence intensity as an index representing reactivity, noise can be removed, so that more accurate detection and / or analysis can be performed.

[0065] (e) Selection of combination of labeled molecule and binding molecule: Processing unit 12 In this technology, based on the calculated reactivity between the target molecule and the binding molecule, a preferable combination of the labeled molecule and the binding molecule for the target molecule can be selected.

[0066] For example, when using the signal / background ratio of fluorescence intensity as an index representing reactivity, a combination of a labeled molecule and a binding molecule with a signal / background ratio of fluorescence intensity equal to or higher than a threshold value can be selected. By doing so, all labeled binding molecules can be detected. Also, a combination of a labeled molecule and a binding molecule with the maximum total signal / background ratio of fluorescence intensity can be selected. By doing so, since the signal / background ratio becomes high, detection becomes easy. Further, by combining these, a combination of a labeled molecule and a binding molecule can be selected.

[0067] Furthermore, for example, a combination of a labeled molecule and a binding molecule with the maximum total difference between the signal of fluorescence intensity and the background of fluorescence intensity can also be selected. By doing so, a signal higher than the background can be obtained, so that detection becomes easy.

[0068] Also, it is possible to select a combination of a labeled molecule and a binding molecule based on the magnitude of the signal of fluorescence intensity. By doing so, since the autofluorescence of the sample serving as the background tends to exhibit high luminance on the short-wavelength side, all signal / background ratios can be increased.

[0069] Furthermore, it is also possible to select combinations of labeled molecules and binding molecules such that, starting from binding molecules with small values of the first signal group, labels with high fluorescence intensity signals are assigned in order. By doing so, it is possible to set advantageous conditions for detection in order starting from binding molecules that are difficult to detect, and it is possible to eliminate combinations that are below the detection limit.

[0070] In addition, it is also possible to select combinations of labeled molecules and binding molecules such that, starting from binding molecules with large values of the first signal group, labels with short detection wavelengths are assigned in order. At this time, when there are multiple candidates, it is possible to select a combination with a higher index calculation value.

[0071] (6) Evaluation unit 13 The information processing apparatus 1 according to the present technology can include an evaluation unit 13 that estimates the significance of binding molecules and / or labeled molecules with respect to target molecules based on image information. Specifically, in the evaluation unit 13, for example, depending on the types of binding molecules and labeled molecules, it is determined whether the luminance should be increased or not, etc., and the method of setting the threshold is considered. When the signal is weak, the background is lowered to make it easier to pick up the target signal. Thus, the significance of binding molecules and / or labeled molecules with respect to target molecules is estimated.

[0072] (7) Output unit 14 The information processing apparatus 1 according to the present technology can include an output unit 14 that outputs various types of information. In the output unit 14, in addition to various types of information related to the calculation of reactivity such as the information processed by the processing unit 12, various signals, and various thresholds, all data such as various types of information related to the detection performed for the calculation of reactivity can be output to the outside.

[0073] Specifically, the output unit 14 can output the matrix of the combination of the binding molecule and the labeling molecule calculated above, and the combination of the binding molecule and the labeling molecule selected above. Further, the output unit 14 can also output the significance of the binding molecule and / or the labeling molecule with respect to the target molecule estimated by the evaluation unit 13 above.

[0074] (8) Presentation unit 15 Based on the output reactivity, the presentation unit 15 presents support information on the combination of the binding molecule and the labeling molecule to the user. Conventionally, the selection of the combination of the labeling molecule and the binding molecule for the target molecule was performed by the user himself / herself, so the combination may not be appropriate, and even if the user is experienced, it is a very laborious task. However, according to the present technology, based on the reactivity calculated by the processing unit 12 and output by the output unit 14, for example, according to the type of the target molecule for detection or analysis and the state of the sample, etc., the optimal combination of the labeling molecule and the binding molecule is presented, so high-precision detection can be performed regardless of the user's experience value.

[0075] In the present technology, the presentation unit 15 is not essential, and it is also possible to automatically label the target molecule by a labeling device or the like without passing through the user. For example, the combination of the binding molecule and the labeling molecule output by the output unit 14 is directly output to various labeling devices, and based on the combination of the binding molecule and the labeling molecule acquired by the various labeling devices, the various labeling devices can automatically label the target molecule using the optimal combination of the binding molecule and the labeling molecule.

[0076] (9) Storage unit 16 The information processing apparatus 1 according to the present technology can be provided with a storage unit 16 for storing various information. The storage unit 16 can accumulate and store all kinds of data such as the information processed by the processing unit 12, various signals, various thresholds, etc., various information related to the calculation of reactivity, and other various information related to the detection performed for the calculation of reactivity.

[0077] The memory unit 16 is not essential in the information processing apparatus 1 according to the present technology. It is also possible to output each piece of information from the output unit 14 to the outside of the apparatus and store it in an external storage device 21 as described later. The storage device 21 can also be provided in a cloud environment and can be connected to the information processing apparatus 1 according to the present technology via a network. In this case, it is also possible to share various information stored in the storage device 21 on the cloud among a plurality of users.

[0078] Based on the information output by the output unit 14, the detection data detected by external detection devices 41A to 41D, etc. as shown in FIG. 7 described later, and the detection data collected from other samples, etc., a database can be constructed in the memory unit 16 and the external storage device 21. In this case, in the processing unit 12, it is also possible to perform various processes with reference to the database. Specifically, it is also possible to refer to a database storing a first signal group, a second signal group, a third signal, a background, and various calculated data. For example, by referring to the data implemented in the past, the detection data detected by external detection devices 41A to 41D, etc., and the detection data collected from other samples, etc., it is possible to calculate the reactivity between the target molecule and the binding molecule for each of the types of the labeled molecule and the binding molecule, and create a matrix of the combination of the binding molecule and the labeled molecule without the user actually performing the measurement.

[0079] Also, by calculating a correction coefficient using various signals aggregated in the database, the accuracy of the correction coefficient can be improved as the number of data increases.

[0080] Furthermore, a database storing a matrix of the combination of the binding molecule and the labeled molecule By referring to it and verifying the results of detecting and / or analyzing the target molecule using the combination of the binding molecule and the labeled molecule, the recommended accuracy of the recommended combination can also be improved.

[0081] (10) Display unit 17 The information processing apparatus 1 according to the present technology can include a display unit 17 that displays various information output by the output unit 14. As the display unit 17, for example, a general display device such as a display or a printer can be used.

[0082] (11) User interface 18 The information processing apparatus 1 according to the present technology can further include a user interface 18 for a user to operate. The user can access and control each unit through the user interface 18.

[0083] In the present technology, the user interface 18 is not essential, and an external operation device may be connected. As the user interface 18, for example, a mouse, a keyboard, or the like can be used.

[0084] <2. Information processing system 2> FIG. 3 is a conceptual diagram showing an example of the information processing system 2 according to the present technology. The information processing system 2 according to the present technology includes the information processing apparatus 1 of the present technology described above, and a storage device 21 that stores the information calculated by the information processing apparatus 1. Further, the information processing system 2 according to the present technology can include an evaluation device 24, a display device 22, a user interface 23, etc. as necessary. Since the details of the information processing apparatus 1 are the same as those of the information processing apparatus 1 of the present technology described above, the description is omitted here. Also, regarding the details of the evaluation device 24, the storage device 21, the display device 22, and the user interface 23, they are the same as the details of the evaluation unit 13, the storage unit 16, the display unit 17, and the user interface 18 of the information processing apparatus 1 of the present technology described above, respectively, so the description is omitted here.

[0085] <3. Biological sample detection device 3, biological sample detection system 4> FIG. 4 is a block diagram showing an example of the biological sample detection device 3 according to the present technology. The biological sample detection device 3 according to the present technology includes a detection unit 31, a signal acquisition unit 11, a processing unit 12, and an output unit 14. Further, the biological sample detection device 3 according to the present technology may be provided with a labeling unit 32, an evaluation unit 13, a presentation unit 15, a storage unit 16, a display unit 17, a user interface 18, an analysis unit 33, etc., as necessary. The details of the signal acquisition unit 11, the processing unit 12, the output unit 14, the evaluation unit 13, the presentation unit 15, the storage unit 16, the display unit 17, and the user interface 18 are the same as those of the signal acquisition unit 11, the processing unit 12, the output unit 14, the evaluation unit 13, the presentation unit 15, the storage unit 16, the display unit 17, and the user interface 18 of the information processing device 1 described above, so the description is omitted here.

[0086] FIG. 5 is a conceptual diagram showing an example of the biological sample detection system 4 according to the present technology. The biological sample detection system 4 according to the present technology includes a detection device 41 and the information processing device 1 according to the present technology described above. Further, the biological sample detection system 4 according to the present technology may be provided with a labeling unit 32 or a labeling device 42, an evaluation unit 13 or an evaluation device 24, a storage unit 16 or a storage device 21, a display device 22, a user interface 23, an analysis unit 33 or an analysis device 43, etc., as necessary. The details of the information processing device 1 are the same as those of the information processing device 1 described above, so the description is omitted here. Also, the details of the evaluation unit 13 or the evaluation device 24, the storage unit 16 or the storage device 21, the display device 22, and the user interface 23 are the same as those of the evaluation unit 13, the storage unit 16, the display unit 17, and the user interface 18 of the information processing device 1 of the present technology described above, so the description is omitted here.

[0087] FIG. 6 is a conceptual diagram showing an example different from FIG. 5 of the biological sample detection system 4 according to the present technology. The biological sample detection system 4 according to the present technology includes a detection device 41 and has a computer program described later.

[0088] (1) Detection unit 31, detection device 41 In the detection unit 31 and the detection device 41, a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule is detected. The combination of the labeling molecule and the binding molecule for labeling the target molecule can be selected based on the reactivity output from the output unit 14.

[0089] As the detection unit 31 and the detection device 41 that can be used in the present technology, as long as the signal emitted from the binding molecule can be detected, general detection units and detection devices can be freely used. For example, detection units and detection devices that can be used in analyses such as flow cytometry, microscopic observation, Western blot, various arrays, and ELISA can be mentioned.

[0090] In the biological sample detection system 4 according to the present technology, the information processing device 1 and / or the storage device 21 can be provided in a cloud environment and connected to the detection device 41 via a network. In this case, it is also possible to share various information stored in the storage device 21 on the cloud among a plurality of users. Specifically, for example, as shown in FIG. 7, a plurality of detection devices 41A to 41D are connected to the information processing device 1 and / or the storage device 21 via a network, and the information processing device 1 processes using the signals detected by the plurality of detection devices 41A to 41D, and it is also possible to share the reactivity matrix of the combination of the labeling molecule and the binding molecule output from the information processing device 1 among the plurality of detection devices 41A to 41D.

[0091] (2) Labeling unit 32, labeling device 42 In the labeling unit 32 and the labeling device 42, the target molecule in the biological sample is labeled using a binding molecule labeled with a labeling molecule. In the labeling unit 32 and the labeling device 42, based on the reactivity matrix of the combination of the labeling molecule and the binding molecule output from the output unit 14, the labeling of the target molecule can be performed using the optimal combination of the labeling molecule and the binding molecule. As a result, the accuracy of target molecule detection can be improved.

[0092] Note that the labeling unit 32 and the labeling device 42 are not essential in the biological sample detection device 3 and the biological sample detection system 4 according to the present technology. Based on the reactivity matrix in the combination of the labeled molecule and the binding molecule output from the output unit 14, it is also possible to label the target molecule using an external labeling device or the like.

[0093] (3) Analysis unit 33, analysis device 43 In the analysis unit 33 and the analysis device 43, the sample is analyzed based on the signals detected by the detection unit 31 and the detection device 41. More specifically, based on the signals detected by the detection unit 31 and the detection device 41, it is possible to analyze the type, amount, properties, etc. of the target molecules contained in the sample.

[0094] Note that the analysis unit 33 and the analysis device 43 are not essential in the biological sample detection device 3 and the biological sample detection system 4 according to the present technology. Based on the signals detected by the detection unit 31 and the detection device 41, it is also possible to analyze the characteristics of the target molecules in the sample using an external analysis device or the like. For example, the analysis unit 33 and the analysis device 43 may be implemented by a personal computer or a CPU, and may be stored as a program in a hardware resource provided with a recording medium (for example, a non-volatile memory (USB memory), HDD, CD, etc.), and may be made to function by a personal computer or a CPU. Further, the analysis unit 33 and the analysis device 43 may be connected to each part of the biological sample detection device 3 and the biological sample detection system 4 via a network.

[0095] <4. Computer program> The computer program according to the present technology is a computer program for causing a computer to realize a signal acquisition function for acquiring a signal derived from a sample containing a biological sample, a processing function for calculating the reactivity of a target molecule with each of a plurality of different binding molecules labeled with different labeling molecules based on the signal, and an output function for outputting the reactivity, wherein the signal includes a first signal group and a second signal group.

[0096] The computer program according to the present technology is recorded on a suitable recording medium. Further, the computer program according to the present technology can be stored in a cloud environment or the like and downloaded and used by a user through a network to a personal computer or the like. Note that the signal acquisition function, the processing function, and the output function in the computer program according to the present technology are the same as the respective functions performed by the signal acquisition unit 11, the processing unit 12, and the output unit 14 of the information processing apparatus 1 described above, and thus the description thereof is omitted here.

[0097] <5. Biological Sample Analysis Method> FIG. 8 is a flowchart showing an example of the biological sample analysis method according to the present technology. The biological sample analysis method according to the present technology is a method that at least performs a signal acquisition step S1, a processing step S2, and an output step S3. Further, if necessary, a presentation step S4, an evaluation step, a storage step, a display step, etc., which are not shown, can also be performed. Note that the signal acquisition step S1, the processing step S2, the output step S3, the presentation step S4, the evaluation step, the storage step, and the display step are the same as the methods performed by the signal acquisition unit 11, the processing unit 12, the output unit 14, the presentation unit 15, the evaluation unit 13, the storage unit 16, and the display unit 17 of the information processing apparatus 1 described above, and thus the description thereof is omitted here.

[0098] <6. Application Examples> The present technology can be applied to, for example, a microscope system 5000 or a biological sample analyzer 6100.

[0099] [Microscope System 5000] A configuration example of the microscope system 5000 of the present disclosure is shown in FIG. 9. The microscope system 5000 shown in FIG. 9 includes a microscope device 5100, a control unit 5110, and an information processing unit 5120. The microscope device 5100 includes a light irradiation unit 5101, an optical unit 5102, and a signal acquisition unit 5103. The microscope device 5100 may further include a sample stage 5104 on which a biological sample S is placed. Note that the configuration of the microscope device 5100 is not limited to that shown in FIG. 9. For example, the light irradiation unit 5101 may be present outside the microscope device 5100, and a light source not included in the microscope device 5100 may be used as the light irradiation unit 5101. Further, the light irradiation unit 5101 may be arranged such that the sample stage 5104 is sandwiched between the light irradiation unit 5101 and the optical unit 5102. For example, it may be arranged on the side where the optical unit 5102 is present. The microscope device 5100 may be configured to be able to perform one or more of bright-field observation, phase-contrast observation, differential interference observation, polarization observation, fluorescence observation, and dark-field observation.

[0100] The microscope system 5000 may be configured as a so-called WSI (Whole Slide Imaging) system or a digital pathology imaging system and can be used for pathological diagnosis. Further, the microscope system 5000 may be configured as a fluorescence imaging system, particularly a multi-fluorescence imaging system.

[0101] For example, the microscope system 5000 may be used for intraoperative pathological diagnosis or remote pathological diagnosis. In the intraoperative pathological diagnosis, during the surgery, the microscope device 5100 can acquire data of the biological sample S obtained from the subject of the surgery and transmit the data to the information processing unit 5120. In the remote pathological diagnosis, the microscope device 5100 can transmit the acquired data of the biological sample S to the information processing unit 5120 existing at a location (such as another room or building) away from the microscope device 5100. Then, in these diagnoses, the information processing unit 5120 receives and outputs the data. Based on the output data, the user of the information processing unit 5120 can perform a pathological diagnosis.

[0102] (Biological sample S) The biological sample S may be a sample containing biological components. The biological components may be biological tissues, cells, liquid components of the living body (such as blood and urine), cultures, or living cells (such as cardiomyocytes, neurons, and fertilized eggs).

[0103] The biological sample S may be a solid, and may be a specimen fixed with a fixing reagent such as paraffin or a solid formed by freezing. The biological sample S may be a section of the solid. As a specific example of the biological sample S, a section of a biopsy sample can be cited.

[0104] The biological sample S may be one subjected to treatments such as staining or labeling. The treatment may be staining for showing the form of biological components or for showing substances (such as surface antigens) possessed by biological components, and examples include HE (Hematoxylin-Eosin) staining and immunohistochemistry staining. The biological sample S may be one subjected to the treatment with one or more reagents, and the reagent may be a fluorescent dye, a chromogenic reagent, a fluorescent protein, or a fluorescently labeled antibody.

[0105] The specimen may be prepared for the purpose of pathological diagnosis or clinical examination from a tissue sample. Further, the specimen is not limited to the human body and may be derived from animals, plants, or other materials. The specimen differs in properties depending on the type of tissue used (such as organs or cells), the type of disease targeted, the attributes of the subject (such as age, gender, blood type, or race), or the lifestyle of the subject (such as diet, exercise habits, or smoking habits). The specimen may be managed with distinguishable identification information (such as a barcode or a QR code (registered trademark)) attached to each specimen.

[0106] (Light irradiation unit 5101) The light irradiation unit 5101 is a light source for illuminating the biological sample S and an optical unit that guides the light irradiated from the light source to the specimen. The light source can irradiate the biological sample S with visible light, ultraviolet light, infrared light, or a combination thereof. The light source may be one or more of a halogen light source, a laser light source, an LED light source, a mercury light source, and a xenon light source. The type and / or wavelength of the light source in fluorescence observation may be plural and may be appropriately selected by those skilled in the art. The light irradiation unit 5101 may have a transmission type, a reflection type, or an epi-illumination type (coaxial epi-illumination type or side illumination type) configuration.

[0107] (Optical unit 5102) The optical unit 5102 is configured to guide the light from the biological sample S to the signal acquisition unit 5103. The optical unit 5102 may be configured to enable the microscope device 5100 to observe or image the biological sample S.

[0108] The optical unit 5102 may include an objective lens. The type of the objective lens may be appropriately selected by those skilled in the art according to the observation method. Further, the optical unit 5102 may include a relay lens for relaying the image magnified by the objective lens to the signal acquisition unit 5103. The optical unit 5102 may further include optical components other than the objective lens and the relay lens, an eyepiece lens, a phase plate, a condenser lens, etc.

[0109] In addition, the optical unit 5102 may further include a wavelength separation unit configured to separate light having a predetermined wavelength from the light from the biological sample S. The wavelength separation unit may be configured to selectively allow light of a predetermined wavelength or wavelength range to reach the signal acquisition unit 5103. The wavelength separation unit may include, for example, one or more of a filter that selectively transmits light, a polarizing plate, a prism (Wollaston prism), and a diffraction grating. The optical components included in the wavelength separation unit may be arranged, for example, on the optical path from the objective lens to the signal acquisition unit 5103. The wavelength separation unit is provided in the microscope device 5100 particularly when fluorescence observation is performed, especially when an excitation light irradiation unit is included. The wavelength separation unit may be configured to separate fluorescent lights from each other or to separate white light and fluorescence.

[0110] (Signal acquisition unit 5103) The signal acquisition unit 5103 can be configured to receive light from the biological sample S and convert the light into an electrical signal, particularly a digital electrical signal. The signal acquisition unit 5103 may be configured to acquire data regarding the biological sample S based on the electrical signal. The signal acquisition unit 5103 may be configured to acquire data of an image (picture, particularly a still image, a time-lapse image, or a moving image) of the biological sample S, and particularly may be configured to acquire data of an image enlarged by the optical unit 5102. The signal acquisition unit 5103 includes one or more image sensors, such as CMOS or CCD, having a plurality of pixels arranged in a one-dimensional or two-dimensional array. The signal acquisition unit 5103 may include an image sensor for acquiring a low-resolution image and an image sensor for acquiring a high-resolution image, or may include a sensing image sensor for AF or the like and an image output sensor for observation or the like. The image sensor may include, in addition to the plurality of pixels, a signal processing unit (including one or two or more of a CPU, a DSP, and a memory) that performs signal processing using pixel signals from each pixel, and an output control unit that controls the output of image data generated from the pixel signals and processing data generated by the signal processing unit. The image sensor including the plurality of pixels, the signal processing unit, and the output control unit may preferably be configured as a one-chip semiconductor device.

[0111] Note that the microscope system 5000 may further include an event detection sensor. The event detection sensor may include pixels that photoelectrically convert incident light and may be configured to detect, as an event, that the luminance change of the pixels exceeds a predetermined threshold value. The event detection sensor may particularly be an asynchronous type.

[0112] (Control unit 5110) The control unit 5110 controls the imaging by the microscope device 5100. For imaging control, the control unit 5110 can drive the movement of the optical unit 5102 and / or the sample stage unit 5104 to adjust the positional relationship between the optical unit 5102 and the sample stage unit 5104. The control unit 5110 can move the optical unit 5102 and / or the sample stage unit 5104 in a direction approaching or separating from each other (for example, the optical axis direction of the objective lens). Also, the control unit 5110 may move the optical unit 5102 and / or the sample stage unit 5104 in any direction in a plane perpendicular to the optical axis direction. For imaging control, the control unit 5110 may control the light irradiation unit 5101 and / or the signal acquisition unit 5103.

[0113] (Sample stage unit 5104) The sample stage unit 5104 may be configured such that the position of the biological sample S on the sample stage unit 5104 can be fixed, and it may be a so-called stage. The sample stage unit 5104 may be configured to be able to move the position of the biological sample S in the optical axis direction of the objective lens and / or in a direction perpendicular to the optical axis direction.

[0114] (Information processing unit 5120) The information processing unit 5120 can acquire data (such as imaging data) acquired by the microscope device 5100 from the microscope device 5100. The information processing unit 5120 can execute image processing on the imaging data. The image processing may include unmixing processing, particularly spectral unmixing processing. The unmixing processing may include processing for extracting data of light components of a predetermined wavelength or wavelength range from the imaging data to generate image data, or processing for removing data of light components of a predetermined wavelength or wavelength range from the imaging data. Also, the image processing may include autofluorescence separation processing for separating the autofluorescence component and the dye component of the tissue section, and fluorescence separation processing for separating wavelengths between dyes having different fluorescence wavelengths from each other. In the autofluorescence separation processing, processing for removing the autofluorescence component from the image information of the other sample using the autofluorescence signal extracted from one of the plurality of samples that are the same or have similar properties may be performed.

[0115] The information processing unit 5120 may transmit data for imaging control to the control unit 5110, and the control unit 5110 that has received the data may control the imaging by the microscope device 5100 according to the data.

[0116] The information processing unit 5120 may be configured as an information processing device such as a general-purpose computer, and may include a CPU, a RAM, and a ROM. The information processing unit 5120 may be included in the housing of the microscope device 5100, or may be outside the housing. Also, various processes or functions by the information processing unit 5120 may be realized by a server computer or a cloud connected via a network.

[0117] The imaging method of the biological sample S by the microscope device 5100 may be appropriately selected by those skilled in the art according to the type of the biological sample S and the purpose of imaging, etc. Examples of the imaging method will be described below.

[0118] One example of the imaging method is as follows. First, the microscope device 5100 can identify an imaging target area. The imaging target area may be identified so as to cover the entire area where the biological sample S exists, or may be identified so as to cover the target part (the part where the target tissue section, target cell, or target lesion exists) of the biological sample S. Next, the microscope device 5100 divides the imaging target area into a plurality of divided areas of a predetermined size, and the microscope device 5100 sequentially images each divided area. Thereby, images of each divided area are acquired.

[0119] As shown in FIG. 10, the microscope device 5100 identifies an imaging target area R that covers the entire biological sample S. Then, the microscope device 5100 divides the imaging target area R into 16 divided areas. Then, the microscope device 5100 images the divided area R1, and then may image any area included in the imaging target area R, such as an area adjacent to the divided area R1. And the imaging of the divided areas is performed until there are no non-imaged divided areas. Note that areas other than the imaging target area R may also be imaged based on the imaging image information of the divided areas.

[0120] In order to image the next divided region after imaging a certain divided region, the positional relationship between the microscope apparatus 5100 and the sample stage 5104 is adjusted. The adjustment may be performed by moving the microscope apparatus 5100, moving the sample stage 5104, or moving both of them. In this example, the imaging device that images each divided region may be a two-dimensional imaging element (area sensor) or a one-dimensional imaging element (line sensor). The signal acquisition unit 5103 may image each divided region via the optical unit 5102. Also, the imaging of each divided region may be continuously performed while moving the microscope apparatus 5100 and / or the sample stage 5104, or the movement of the microscope apparatus 5100 and / or the sample stage 5104 may be stopped during the imaging of each divided region. The imaging target region may be divided such that a part of each divided region overlaps, or the imaging target region may be divided so as not to overlap. Each divided region may be imaged a plurality of times while changing imaging conditions such as the focal length and / or the exposure time.

[0121] Further, the information processing apparatus can generate image data of a wider region by stitching a plurality of adjacent divided regions. By performing the stitching process over the entire imaging target region, an image of a wider region can be obtained for the imaging target region. Also, image data with a lower resolution can be generated from the image of the divided region or the image on which the stitching process has been performed.

[0122] Other examples of imaging methods are as follows. First, the microscope apparatus 5100 can identify an imaging target area. The imaging target area may be identified so as to cover the entire area where the biological sample S exists, or may be identified so as to cover the target part (the part where the target tissue section or target cells exist) of the biological sample S. Next, the microscope apparatus 5100 images a partial area of the imaging target area (also referred to as a "divided scan area") by scanning it in one direction (also referred to as a "scan direction") in a plane perpendicular to the optical axis. When the scan of the divided scan area is completed, next, the divided scan area adjacent to the scan area is scanned. These scan operations are repeated until the entire imaging target area is imaged.

[0123] As shown in FIG. 11, the microscope apparatus 5100 identifies the area (gray part) where the tissue section exists in the biological sample S as the imaging target area Sa. Then, the microscope apparatus 5100 scans the divided scan area Rs in the Y-axis direction within the imaging target area Sa. When the scan of the divided scan area Rs is completed, the microscope apparatus 5100 then scans the adjacent divided scan area in the X-axis direction. This operation is repeated until the scan is completed for all of the imaging target area Sa.

[0124] For scanning each divided scan region and for imaging the next divided scan region after imaging a certain divided scan region, the positional relationship between the microscope apparatus 5100 and the sample stage 5104 is adjusted. The adjustment may be performed by moving the microscope apparatus 5100, moving the sample stage 5104, or moving both of them. In this example, the imaging device for imaging each divided scan region may be a one-dimensional imaging element (line sensor) or a two-dimensional imaging element (area sensor). The signal acquisition unit 5103 may image each divided region through the magnifying optical system. Also, the imaging of each divided scan region may be continuously performed while moving the microscope apparatus 5100 and / or the sample stage 5104. The imaging target region may be divided such that a part of each divided scan region overlaps, or the imaging target region may be divided so as not to overlap. Each divided scan region may be imaged a plurality of times while changing imaging conditions such as the focal length and / or the exposure time.

[0125] Further, the information processing apparatus can generate image data of a wider region by stitching a plurality of adjacent divided scan regions. By performing the stitching process over the entire imaging target region, an image of a wider region can be acquired for the imaging target region. Also, image data with a lower resolution can be generated from the image of the divided scan region or the image subjected to the stitching process.

[0126] [Biological sample analyzer 6100] A configuration example of the biological sample analyzer 6100 of the present technology is shown in FIG. 12. The biological sample analyzer 6100 shown in FIG. 12 includes a light irradiation unit 6101 that irradiates light onto the biological sample S flowing through the flow path C, a detection unit 6102 that detects the light generated by irradiating the biological sample S with light, and an information processing unit 6103 that processes information regarding the light detected by the detection unit 6102. Examples of the biological sample analyzer 6100 include a flow cytometer and an imaging cytometer. The biological sample analyzer 6100 may include a sorting unit 6104 that sorts a specific biological particle P in the biological sample S. An example of the biological sample analyzer 6100 including the sorting unit is a cell sorter.

[0127] (Biological sample S) The biological sample S may be a liquid sample containing biological particles P. The biological particles P are, for example, cells or acellular biological particles. The cells may be live cells, and more specific examples include blood cells such as red blood cells and white blood cells, and germ cells such as sperm and fertilized eggs. The cells may also be directly collected from a specimen such as whole blood, or may be cultured cells obtained after culturing. Examples of the acellular biological particles include extracellular vesicles, particularly exosomes and microvesicles. The biological particles P may be labeled with one or more labeling substances (for example, dyes (particularly fluorescent dyes) and fluorescent dye-labeled antibodies). Note that particles other than the biological particles P may be analyzed by the biological sample analyzer of the present disclosure, and beads or the like may be analyzed for calibration or the like.

[0128] (Flow path C) The flow path C is configured such that the biological sample S flows therethrough. In particular, the flow path C can be configured such that a flow in which the biological particles P contained in the biological sample S are arranged in a substantially single row is formed. The flow path structure including the flow path C may be designed such that a laminar flow is formed. In particular, the flow path structure is designed such that a laminar flow in which the flow of the biological sample S (sample flow) is surrounded by the flow of the sheath fluid is formed. The design of the flow path structure may be appropriately selected by those skilled in the art, and known ones may be adopted. The flow path C may be formed in a flow path structure such as a microchip (a chip having a flow path on the order of micrometers) or a flow cell. The width of the flow path C is 1 mm or less, and in particular, it may be 10 μm or more and 1 mm or less. The flow path C and the flow path structure including the same may be formed of a material such as plastic or glass.

[0129] The biological sample analyzer 6100 of the present technology is configured such that the light from the light irradiation unit 6101 irradiates the biological sample S flowing in the flow path C, particularly the biological particles P in the biological sample S. The biological sample analyzer 6100 of the present technology may be configured such that the light irradiation point (interrogation point) for the biological sample S is in the flow path structure in which the flow path C is formed, or the light irradiation point may be outside the flow path structure. As an example of the former, a configuration in which the light irradiates the flow path C in a microchip or a flow cell can be given. In the latter case, the light may irradiate the biological particles P after they exit from the flow path structure (particularly its nozzle portion), and for example, a flow cytometer of the Jet in Air type can be given.

[0130] (Light irradiation unit 6101) The light irradiation unit 6101 includes a light source unit that emits light and a light guiding optical system that guides the light to the irradiation point. The light source unit includes one or more light sources. The type of the light source is, for example, a laser light source or an LED. The wavelength of the light emitted from each light source may be any wavelength of ultraviolet light, visible light, or infrared light. The light guiding optical system includes optical components such as, for example, a group of beam splitters, a group of mirrors, or an optical fiber. Further, the light guiding optical system may include a group of lenses for condensing light, for example, an objective lens. The number of irradiation points where the light intersects the biological sample S may be one or more. The light irradiation unit 6101 may be configured to condense the light irradiated from one or a plurality of different light sources onto one irradiation point.

[0131] (Detection unit 6102) The detection unit 6102 includes at least one photodetector that detects the light generated by irradiating the biological particles P with light. The light to be detected is, for example, fluorescence or scattered light (for example, any one or more of forward scattered light, backward scattered light, and lateral scattered light). Each photodetector includes one or more light receiving elements, for example, has a light receiving element array. Each photodetector may include, as the light receiving element, one or a plurality of PMTs (photomultiplier tubes) and / or photodiodes such as APDs and MPPCs. The photodetector includes, for example, a PMT array in which a plurality of PMTs are arranged in a one-dimensional direction. Further, the detection unit 6102 may include an imaging device such as a CCD or a CMOS. The detection unit 6102 can acquire an image of the biological particles P (for example, a bright field image, a dark field image, and a fluorescence image, etc.) by the imaging device.

[0132] The detection unit 6102 includes a detection optical system that causes light of a predetermined detection wavelength to reach the corresponding photodetector. The detection optical system includes a spectroscopic unit such as a prism or a diffraction grating, or a wavelength separation unit such as a dichroic mirror or an optical filter. The detection optical system is configured to, for example, split the light generated by irradiating the biological particle P with light, and have the split light detected by a plurality of photodetectors that is more than the number of fluorescent dyes with which the biological particle P is labeled. A flow cytometer including such a detection optical system is called a spectral flow cytometer. Further, the detection optical system is configured to, for example, separate light corresponding to the fluorescence wavelength range of a specific fluorescent dye from the light generated by irradiating the biological particle P with light, and cause the separated light to be detected by the corresponding photodetector.

[0133] Further, the detection unit 6102 may include a signal processing unit that converts the electrical signal obtained by the photodetector into a digital signal. The signal processing unit may include an A / D converter as a device that performs the conversion. The digital signal obtained by the conversion by the signal processing unit may be transmitted to the information processing unit 6103. The digital signal may be handled by the information processing unit 6103 as data related to light (hereinafter also referred to as "light data"). The light data may be, for example, light data including fluorescence data. More specifically, the light data may be light intensity data, and the light intensity may be light intensity data of light including fluorescence (which may include feature amounts such as Area, Height, Width, etc.).

[0134] (Information processing unit 6103) The information processing unit 6103 includes, for example, a processing unit that executes processing of various data (for example, light data) and a storage unit that stores various data. When the processing unit acquires light data corresponding to a fluorescent dye from the detection unit 6102, it may perform fluorescence spillover correction (compensation processing) on the light intensity data. Further, in the case of a spectral flow cytometer, the information processing unit 6103 executes fluorescence separation processing on the light data and acquires light intensity data corresponding to the fluorescent dye.

[0135] The fluorescence separation process may be performed according to, for example, the unmixing method described in Japanese Patent Application Laid-Open No. 2011-232259. When the detection unit 6102 includes an imaging device, the information processing unit 6103 may acquire the morphological information of the biological particles P based on the image acquired by the imaging device. The storage unit may be configured to store the acquired optical data. The storage unit may further be configured to store the spectral reference data used in the unmixing process.

[0136] When the biological sample analyzer 6100 includes a fractionation unit 6104 described later, the information processing unit 6103 can execute a determination as to whether to fractionate the biological particles P based on the optical data and / or the morphological information. Then, the information processing unit 6103 controls the fractionation unit 6104 based on the result of the determination, and the fractionation of the biological particles P by the fractionation unit 6104 can be performed.

[0137] The information processing unit 6103 may be configured to output various data (such as optical data and images). For example, the information processing unit 6103 can output various data (such as two-dimensional plots, spectral plots, etc.) generated based on the optical data. Also, the information processing unit 6103 may be configured to receive inputs of various data, and for example, receive gating processing on a plot by a user. The information processing unit 6103 may include an output unit (such as a display) or an input unit (such as a keyboard) for executing the output or the input.

[0138] The information processing unit 6103 may be configured as a general-purpose computer, for example, as an information processing device including a CPU, a RAM, and a ROM. The information processing unit 6103 may be included in the housing in which the light irradiation unit 6101 and the detection unit 6102 are provided, or may be outside the housing. Also, various processes or functions by the information processing unit 6103 may be realized by a server computer or a cloud connected via a network.

[0139] (Fractionation unit 6104) The fractionation unit 6104 performs fractionation of the biological particles P according to the determination result by the information processing unit 6103. The fractionation method may be a method of generating droplets containing the biological particles P by vibration, applying a charge to the droplets to be fractionated, and controlling the traveling direction of the droplets with electrodes. The fractionation method may also be a method of controlling the traveling direction of the biological particles P in the flow channel structure and performing fractionation. The flow channel structure is provided with a control mechanism by, for example, pressure (injection or suction) or charge. As an example of the flow channel structure, there is a flow channel structure in which the flow channel C branches into a recovery flow channel and a waste liquid flow channel downstream thereof, and a chip in which specific biological particles P are recovered into the recovery flow channel (for example, the chip described in Japanese Patent Application Laid-Open No. 2020-76736) can be mentioned.

Example

[0140] Hereinafter, the present invention will be described in more detail based on examples. It should be noted that the examples described below show an example of a typical example of the present invention, and the scope of the present invention is not construed narrowly thereby.

[0141] <Experimental Example 1> In Experimental Example 1, a second signal group was obtained when the same type of binding molecules labeled with different labeling molecules were each reacted with a target molecule.

[0142] Specifically, a paraffin-embedded tissue section of human breast cancer was used as an example of a target molecule, AF488, AF647, AF700 of the fluorescent dye AlexaFluor (registered trademark) series, and PE (Phycoerythrin) were used as examples of labeling molecules, and anti-Pan-Cytokeratin antibody (Clone: AE1 / AE3) was used as an example of a binding molecule for fluorescence immunostaining.

[0143] The results of the fluorescence immunostaining are shown in FIG. 13. As shown in FIG. 13, it was confirmed that when the same type of binding molecules labeled with different labeling molecules were each reacted with the target molecule, it affected the reactivity.

[0144] <Experimental Example 2> In Experimental Example 2, a first signal group and a second signal group were acquired, and the reactivity of each of a target molecule with a plurality of different binding molecules labeled with different labeling molecules was calculated.

[0145] Specifically, a formalin-fixed paraffin-embedded tissue section of human tonsil was used as an example of a target molecule, AF488, AF555, AF594, and AF647 of the fluorescent dye AlexaFluor (registered trademark) series were used as examples of labeling molecules, and CD3 antibody, CD5 antibody, and CD7 antibody were used as examples of binding molecules for fluorescence immunostaining.

[0146] (1) Acquisition of the first signal group Imaging images were obtained when a plurality of different binding molecules CD3 antibody, CD5 antibody, and CD7 antibody labeled with the same type of labeling molecule AF647 were each reacted with a target molecule. In addition, imaging images were obtained when the binding molecule CD3 antibody labeled with AF488, the binding molecule CD3 antibody labeled with AF555, and the binding molecule CD5 antibody labeled with AF594 were each reacted with a target molecule. The number of each binding molecule was calculated from the obtained imaging images. The calculated number of binding molecules is shown in Table 4 below.

[0147]

Table 4

[0148] (2) Acquisition of the second signal group Imaging images were obtained when the same type of binding molecule CD3 antibody labeled with different labeling molecules AF488, AF555, and AF647 was each reacted with a target molecule. In addition, imaging images were obtained when the binding molecule CD3 antibody labeled with AF488, the binding molecule CD3 antibody labeled with AF555, and the binding molecule CD5 antibody labeled with AF594 were each reacted with a target molecule. The fluorescence intensity / autofluorescence ratio of each was calculated from the obtained imaging images. The calculated fluorescence intensity / autofluorescence ratio is shown in Table 5 below.

[0149] [Table 5]

[0150] (3) Calculation of reactivity index Using the numerical values in Table 4 and Table 5, the reactivity index was calculated. In this experimental example, the fluorescence intensity / auto-fluorescence ratio was used as the reactivity index. Specifically, the reactivity index was calculated using the following formula. Reactivity index calculated value = (number of antibodies of the same type of label for the binding molecule to be calculated / number of antibodies of the same type of label for the reference binding molecule) × [fluorescence intensity / auto-fluorescence ratio] of the reference binding molecule for the fluorescent dye to be calculated

[0151] For example, the reactivity index (fluorescence intensity / auto-fluorescence ratio) of CD7 labeled with AF488 was calculated as follows. Number of antibodies of the same type of label for the binding molecule to be calculated: number of antibodies of AF647CD7 = 29.5 (Table 4) Number of antibodies of the same type of label for the reference binding molecule: number of antibodies of AF647CD3 = 3.40 (Table 4) [Fluorescence intensity / auto-fluorescence ratio] of the reference binding molecule for the fluorescent dye to be calculated: [fluorescence intensity / auto-fluorescence ratio] of AF488CD3 = 0.739 (29.5 / 3.40) × 0.739 ≈ 6.42

[0152] (4) Calculation results of reactivity index The calculation results of the reactivity index are shown in Table 6 below.

[0153] [Table 6]

[0154] (5) Selection of combinations of labeled molecules and binding molecules Based on the calculated reactivity index, combinations of labeled molecules and binding molecules were selected. In Experimental Example 2, as an example of selection, from the binding molecules with small values in the first signal group, labels with the highest signals were sequentially assigned for selection.

[0155] Specifically, first, in Table 4, the minimum value of the first signal in AF647 is 1.92 of AF647CD5, and the maximum value of the calculated reactivity index of CD5 is 2.58 of AF647CD5. Therefore, the label AF647 was assigned to the binding molecule CD5.

[0156] In Table 4, the second smallest value of the first signal in AF647 is 3.40 of AF647CD3, and the maximum value of the calculated reactivity index of CD3 is 4.58 of AF647CD3. However, since AF647 has already been assigned to CD5, 2.41 of AF555CD3, which shows the next highest value, was selected, and the label AF555 was assigned to the binding molecule CD3.

[0157] In Table 4, the second smallest value of the first signal in AF647 is 29.5 of AF647CD7, and the maximum value of the calculated reactivity index of CD7 is 39.8 of AF647CD7. Next is 20.9 of AF555CD7. However, since both have already been assigned, 6.42 of AF488CD7, which shows the next highest value, was selected, and AF488 was assigned to CD7.

[0158] From the above, AF488CD7, AF555CD3, and AF647CD5 were selected as the combinations of the labeling molecule and the binding molecule.

[0159] (6) Staining confirmation Formalin-fixed paraffin-embedded tissue sections of human tonsils were subjected to fluorescence immunostaining using the combinations of the labeling molecule and the binding molecule selected above. Images of each fluorescently labeled antibody in the multiplex staining images are shown in FIGS. 14 to 16. Images of each fluorescent label in non-stained specimens are also shown as negative controls. In addition, for the label AF488, a staining image of AF488CD5, which is a combination not selected because the calculated reactivity index value was low, is also shown for reference.

[0160] (7) Quantification of the number of antibodies The number of each antibody was also calculated from the multiplex staining images using the selected combination of labeled molecules and binding molecules and the images of each fluorescent label in the unstained specimen. The calculation results are shown in Tables 7 and 8 below.

[0161]

Table 7

[0162]

Table 8

[0163] (8) Consideration As shown in FIGS. 14 to 16, when fluorescence immunostaining was performed using the selected combination of labeled molecules and binding molecules, a high value was shown as compared with the negative control, and it was proved that all were detectable. Also, as shown in FIG. 14, AF488CD5, which is a combination not selected because the calculated value of the reactivity index is low, has a lower signal value compared with the selected AF647CD5, and it is difficult to detect. Therefore, it was proved that the combination of AF488CD5 is inappropriate.

[0164] <Experimental Example 3> In Experimental Example 3, a combination of labeled molecules and binding molecules was selected by a method different from that in Experimental Example 2.

[0165] Based on the reactivity index calculated in Experimental Example 2, a combination of labeled molecules and binding molecules was selected. In Experimental Example 3, as an example of the selection, from the binding molecules with large values in the first signal group, fluorescent labels on the short-wavelength side were sequentially assigned for selection. When there were multiple candidates, a combination with a higher calculated value of the panel design index was selected.

[0166] Specifically, first, in Table 4, CD7, CD3, and CD5 are in the order of the largest first signal in AF647, and their values are 29.5, 3.40, and 1.92, respectively. Therefore, AF488 and AF555, which are fluorescent dyes on the short-wavelength side, were assigned in the order of the largest first signal, with CD7 assigned to AF488 and CD3 assigned to AF555. Although it is possible to assign AF594 and AF647 to CD5, since the calculated reactivity index value of AF594CD5 was 0.320 and the calculated reactivity index value of AF647CD5 was 2.58, AF647CD5 with the higher calculated reactivity index value was selected.

[0167] From the above, AF488CD7, AF555CD3, and AF647CD5 were selected as combinations of the labeling molecule and the binding molecule.

[0168] Note that in this technology, the following configurations can also be adopted. (1) A processing unit that calculates the reactivity of each of the target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal from the sample containing the biological sample. The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with the target molecule, and A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with the target molecule, An information processing device including. (2) The information processing device according to (1), which calculates the number of binding molecules and / or the fluorescence intensity labeled on the binding molecule as an index of the reactivity. (3) The information processing device according to (2), wherein the fluorescence intensity is calculated from one or more numerical values selected from excitation efficiency, quantum yield, absorption efficiency, and photolabeling rate (F / P value). (4) The signal includes at least one of a signal, a specific signal / background, and a specific signal / nonspecific signal, and the information processing apparatus according to any one of (1) to (3). (5) In the processing unit, based on a third signal obtained when using a negative control, the background in each detection channel is calculated, and the information processing apparatus according to any one of (1) to (3). (6) In the processing unit, the leakage of the autofluorescence signal and / or the leakage of the signal derived from other labeled molecules are calculated, and the information processing apparatus according to (4). (7) In the processing unit, for a combination of a labeled molecule and a binding molecule that has not actually been measured, the reactivity with the target molecule is calculated, and the information processing apparatus according to any one of (1) to (6). (8) In the processing unit, a combination of a labeled molecule and a binding molecule in which the specific signal / background is equal to or greater than a threshold value is selected, and the information processing apparatus according to any one of (4) to (7). (9) In the processing unit, a combination of a labeled molecule and a binding molecule in which the total of the specific signal / background is maximized is selected, and the information processing apparatus according to any one of (4) to (8). (10) In the processing unit, a combination of a labeled molecule and a binding molecule in which the total of the difference between the signal of the fluorescence intensity and the background of the fluorescence intensity is maximized is selected, and the information processing apparatus according to any one of (2) to (9). (11) In the processing unit, a combination of a labeled molecule and a binding molecule is selected based on the magnitude of the signal of the fluorescence intensity, and the information processing apparatus according to any one of (2) to (10). (12) In the processing unit, a combination of a labeled molecule and a binding molecule is selected such that, in order from the binding molecule with a small value of the first signal group and in order from the labeled molecule with a high fluorescence intensity signal, and the information processing apparatus according to any one of (2) to (10). (13) The information processing apparatus according to any one of (2) to (10), wherein the processing unit selects a combination of a labeled molecule and a binding molecule so as to assign the values of the first signal group in order from the binding molecules having large values and the labels having short detection wavelengths in order. (14) The information processing apparatus according to any one of (1) to (13), further comprising a presentation unit that presents support information on a combination of a binding molecule and a labeled molecule to a user based on the output reactivity. (15) The information processing apparatus according to any one of (1) to (14), further comprising an evaluation unit that estimates the significance of a binding molecule and / or a labeled molecule with respect to a target molecule based on image information. (16) In the information processing apparatus according to any one of (1) to (15), the first signal group and the second signal group are detection amounts normalized using at least one selected from excitation power density, exposure time, and detection device sensitivity. (17) A signal acquisition step of acquiring a signal from a sample containing a biological sample; A processing step of calculating the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeled molecules based on the signal; An output step of outputting the reactivity, and The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeled molecule are each reacted with a target molecule; and A second signal group obtained when the same type of binding molecules labeled with different labeled molecules are each reacted with a target molecule, A biological sample analysis method comprising: (18) A signal acquisition unit that acquires a signal from a sample containing a biological sample; A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeled molecules based on the signal; an output unit that outputs the reactivity, and the signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, and an information processing apparatus including a storage device that stores information calculated by the information processing apparatus, and an information processing system comprising (19) the information processing system according to (18), wherein information stored in the storage device is taken into consideration in the information processing apparatus (20) a signal acquisition unit that acquires a signal from a sample including a biological sample, a processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal, an output unit that outputs the reactivity, and a detection unit that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity, and has the signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, and a biological sample detection device including (21) the biological sample detection device according to (20), further comprising a labeling unit that labels a target molecule with a binding molecule labeled with a labeling molecule selected based on the output reactivity (22) The biological sample detection device according to (20) or (21), further comprising an analysis unit that analyzes the sample based on the signal detected by the detection unit. (23) A signal acquisition unit that acquires a signal derived from a sample containing a biological sample, A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal, An output unit that outputs the reactivity, and having, The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, An information processing apparatus including A detection apparatus that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity, A biological sample detection system comprising (24) A signal acquisition function for acquiring a signal derived from a sample containing a biological sample, A processing function for calculating the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal, An output function for outputting the reactivity, which is a computer program for causing a computer to realize, The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, A computer program including (25) A signal acquisition function for acquiring signals from a sample containing a biological sample, a processing function for calculating the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeling molecules based on the signal, an output function for outputting the reactivity, which is a computer program for causing a computer to implement, wherein the signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, a computer program including, a detection device for detecting a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity, and a biological sample detection system having.

[0169] 1 Information processing apparatus 11 Signal acquisition unit 12 Processing unit 13 Evaluation unit 14 Output unit 15 Presentation unit 16 Storage unit 17 Display unit 18, 23 User interface 2 Information processing system 3 Biological sample detection device 4 Biological sample detection system 21 Storage device 22 Display device 24 Evaluation device 31, 6102 Detection unit 41 Detection device 32 Labeling unit 42 Labeling device 33 Analysis unit 43 Analysis device 5000 Microscope System 5100 Microscope Device 5110 Control Unit 5120, 6103 Information Processing Unit 5101, 6101 Light Irradiation Unit 5102 Optical Unit 5103 Signal Acquisition Unit 5104 Specimen Placement Unit S Biological Sample, Biological Specimen R, Sa Imaging Target Region R1 Division Region Rs Division Scan Region 6100 Biological Specimen Analyzer C Flow Path P Biological Particle 6104 Sampling Unit

Claims

1. A processing unit that calculates the reactivity of a target molecule with a plurality of different binding molecules labeled with different labeling molecules based on signals from a sample containing a biological sample, wherein the signal is a first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule, and is a second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule, An information processing apparatus comprising:

2. The information processing apparatus according to claim 1, wherein the processing unit calculates the number of binding molecules and / or the fluorescence intensity labeled on the binding molecules as an index of the reactivity.

3. The information processing apparatus according to claim 2, wherein the fluorescence intensity is calculated from one or more numerical values selected from excitation efficiency, quantum yield, absorption efficiency, and photolabeling rate (F / P value).

4. The information processing apparatus according to claim 1, wherein the signal includes at least one of a signal, a specific signal / background, and a specific signal / nonspecific signal.

5. The information processing apparatus according to claim 1, wherein the processing unit calculates the background in each detection channel based on a third signal obtained when a negative control is used.

6. The information processing apparatus according to claim 2, wherein the processing unit calculates the leakage of autofluorescence signals and / or the leakage of signals from other labeling molecules.

7. The information processing apparatus according to claim 1, wherein the processing unit calculates the reactivity of a target molecule with a combination of a labeling molecule and a binding molecule that has not actually been measured.

8. The information processing apparatus according to claim 4, wherein the processing unit selects a combination of a labeling molecule and a binding molecule for which the specific signal / background is equal to or greater than a threshold value.

9. The information processing apparatus according to claim 4, wherein the processing unit selects a combination of a labeling molecule and a binding molecule for which the total of the specific signal / background is maximized.

10. The information processing apparatus according to claim 2, wherein the processing unit selects a combination of a labeling molecule and a binding molecule for which the total of the difference between the fluorescence intensity signal and the fluorescence intensity background is maximized.

11. The information processing apparatus according to claim 2, wherein the processing unit selects a combination of a labeled molecule and a binding molecule based on the magnitude of the signal of the fluorescence intensity.

12. The information processing apparatus according to claim 2, wherein the processing unit selects a combination of a labeled molecule and a binding molecule such that, in order from the binding molecules with small values of the first signal group, and in order from the labels with high fluorescence intensity signals.

13. The information processing apparatus according to claim 2, wherein the processing unit selects a combination of a labeled molecule and a binding molecule such that, in order from the binding molecules with large values of the first signal group, and in order from the labels with short detection wavelengths.

14. The information processing apparatus according to claim 1, further comprising a presentation unit that presents, to the user, support information on a combination of a binding molecule and a labeled molecule based on the calculated reactivity.

15. The information processing apparatus according to claim 1, further comprising an evaluation unit that estimates the significance of a binding molecule and / or a labeled molecule with respect to a target molecule based on image information.

16. The information processing apparatus according to claim 1, wherein the first signal group and the second signal group are detection amounts normalized using at least one selected from excitation power density, exposure time, and detection device sensitivity.

17. A signal acquisition step of acquiring a signal from a sample containing a biological sample; A processing step of calculating the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeled molecules based on the signal; An output step of outputting the reactivity, and wherein the signal includes a first signal group acquired when a plurality of different binding molecules labeled with the same type of labeled molecule are each reacted with a target molecule; and a second signal group acquired when a plurality of the same type of binding molecules labeled with different labeled molecules are each reacted with a target molecule. A biological sample analysis method.

18. A signal acquisition unit that acquires a signal from a sample containing a biological sample; A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when using a plurality of different binding molecules labeled with different labeled molecules based on the signal; An output unit that outputs the reactivity A detection unit that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule, the binding molecule being selected based on the output reactivity; The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule; A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule; A biological sample detection device including the above.

19. The biological sample detection device according to claim 18, further comprising a labeling unit that labels a target molecule using a binding molecule labeled with a labeling molecule selected based on the output reactivity.

20. The biological sample detection device according to claim 18, further comprising an analysis unit that analyzes the sample based on the signal detected by the detection unit.

21. A signal acquisition unit that acquires a signal from a sample containing a biological sample; A processing unit that calculates the reactivity of each of a target molecule and a plurality of different binding molecules when a plurality of different binding molecules labeled with different labeling molecules are used based on the signal; An output unit that outputs the reactivity, and The signal is A first signal group obtained when a plurality of different binding molecules labeled with the same type of labeling molecule are each reacted with a target molecule; A second signal group obtained when the same type of binding molecules labeled with different labeling molecules are each reacted with a target molecule; An information processing device including the above, and A detection device that detects a signal emitted from a target molecule labeled with a binding molecule labeled with a labeling molecule selected based on the output reactivity; A biological sample detection system comprising the above.

Images