Measuring device and analysis method

Abstract

To provide a measuring device with which measurement throughput increases.SOLUTION: Provided is a measuring device for analyzing a cell included in a specimen. The measuring device includes: a chamber for preparing a measurement sample in which the cell is dyed by a first and a second fluorescent dye that are included in the reagent supplied from at least one reagent container; a liquid sending unit for sending the reagent from the reagent container to the chamber via a liquid sending tube that is provided between the reagent container and the chamber; a detection unit that acquires a first and a second signal corresponding respectively to the fluorescence of a first wavelength and the fluorescence of a second wavelength radiated from the cell having been dyed with the first and second fluorescent dyes, in accordance with irradiation with light of the measurement sample flowing in a flow cell; and an analysis unit that analyzes the cell on the basis of the first and second signals.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a measurement device and an analysis method. [Background technology]

[0002] Non-Patent Document 1 discloses a flow cytometer that analyzes signals emitted from multiple fluorescent dyes with different fluorescent wavelength bands. Measurements using the flow cytometer disclosed in Non-Patent Document 1 use a reagent (see Non-Patent Document 2) that stains a sample with multiple fluorescent dyes with different wavelength bands. Each fluorescent dye is attached to an antibody, and each antibody stains the sample by binding to the measurement target in the sample. The flow cytometer aspirates antibody reagent from a reagent container using a dispensing probe and a mechanism for moving the dispensing probe, and ejects the antibody reagent into a reaction container that contains a sample having the measurement target. The flow cytometer measures the measurement target that has been mixed with the antibody reagent and stained in the reaction container. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] AQUIOS Tetra System Guide https: / / www.beckmancoulter.com / wsrportal / techdocs?docname=B26364AB.pdf [Non-Patent Document 2] AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panelhttps: / / www.beckman.jp / techdocs / B25337AG / wsr-161331 Summary of the Invention [Problem to be solved by the invention]

[0004] In the flow cytometer disclosed in Non-Patent Document 1, in order to prepare a measurement sample, a reagent is aspirated from a reagent container using a nozzle, the nozzle that has aspirated the reagent is moved to the location of the reaction container, and the reagent is discharged into the reaction container. In this case, since a process is required for aspirating and discharging the reagent, it takes time to prepare the measurement sample (Non-Patent Document 1 states that the measurement throughput is 25 samples per hour).

[0005] An object of the present invention is to provide a measurement device and an analysis method that are capable of implementing measurements using multiple fluorescent dyes with high processing capacity. [Means for solving the problem]

[0006] The measuring device of the present invention is a measuring device for analyzing cells contained in a specimen, and includes: a chamber for preparing a measurement sample in which the cells are stained with first and second fluorescent dyes contained in a reagent supplied from at least one reagent container; a liquid delivery unit for delivering the reagent from the reagent container to the chamber via a liquid delivery tube provided between the reagent container and the chamber; a detection unit for acquiring first and second signals corresponding to the fluorescence of a first wavelength and the fluorescence of a second wavelength, respectively, emitted from the cells stained with the first and second fluorescent dyes in response to irradiation of the measurement sample flowing in a flow cell with light; and an analysis unit for analyzing the cells based on the first and second signals. The analytical method of the present invention is an analytical method for analyzing cells contained in a specimen, which comprises the steps of: delivering a reagent from a reagent container to a chamber via a delivery tube provided between the reagent container and a chamber for mixing the specimen and the reagent to prepare a measurement specimen; preparing the measurement specimen in which the cells are stained with first and second fluorescent dyes contained in the reagent supplied from at least one of the reagent containers; irradiating light onto the measurement specimen flowing through a flow cell; acquiring first and second fluorescent signals corresponding to the fluorescence of a first wavelength and the fluorescence of a second wavelength, respectively, emitted from the cells stained with the first and second fluorescent dyes in response to the irradiation of light; and analyzing the cells based on the first and second fluorescent signals. Effect of the Invention

[0007] According to the present invention, it is possible to provide an analytical device capable of performing measurements using a plurality of fluorescent dyes with high processing capacity. [Brief description of the drawings]

[0008] [Figure 1] 1 is a perspective view showing an analysis system according to a first embodiment of the present invention. [Diagram 2] FIG. 2 is a schematic diagram showing the configuration of a measurement unit according to the first embodiment of the present invention. [Diagram 3] 4 is a flowchart showing the procedure of a measurement sample preparation process performed by the analysis system according to the first embodiment of the present invention. [Figure 4] FIG. 13 is a block diagram showing the configuration of a measurement unit of an analysis system according to a second embodiment of the present invention. [Diagram 5] FIG. 11 is a diagram showing a fluid circuit including a specimen aspirating section, a sample preparing section, and a detecting section in a second embodiment of the present invention. [Figure 6] FIG. 11 is a schematic diagram showing another example of the first sample preparation unit in the second embodiment of the present invention. [Figure 7] FIG. 11 is a configuration explanatory diagram showing an example of an optical system of an FCM detection section in a second embodiment of the present invention. [Figure 8] 13 is a configuration explanatory diagram showing another example of the optical system of the FCM detection section in the second embodiment of the present invention. FIG. [Figure 9] FIG. 1 shows an example of fluorescence emitted from two different fluorescent dyes bleeding into each other. [Figure 10] FIG. 11 is a block diagram showing an example of a configuration of an analysis unit in a second embodiment of the present invention. [Figure 11] FIG. 11 is a perspective view showing another configuration example of the analysis system in the second embodiment of the present invention. [Figure 12] FIG. 11 is a block diagram showing the configuration of another example of the measurement unit in the second embodiment of the present invention. [Figure 13]FIG. 11 is a diagram showing a state in which the cover of a measurement unit in a second embodiment of the present invention is open. [Figure 14] FIG. 11 is a perspective view showing a reagent container holder of a measuring unit in a second embodiment of the present invention. [Figure 15] 15 is a front view showing the reagent container holder shown in FIG. 14. [Figure 16] 15 is a schematic diagram for explaining a reagent container holding portion of the reagent container holder shown in FIG. 14. [Figure 17] 15 is a schematic diagram showing a state in which a reagent container is placed on the reagent container holder shown in FIG. 14. FIG. [Figure 18] 15 is a schematic diagram showing a state in which a reagent container is placed on the reagent container holder shown in FIG. 14. FIG. [Figure 19] 15 is a longitudinal sectional view showing a schematic internal configuration of the reagent container holder shown in FIG. 14. [Figure 20] 20 is a longitudinal sectional view of the reagent container holder shown in FIG. 19, illustrating a state in which the reagent container is set. FIG. [Figure 21] 21 is a vertical sectional view for explaining the state in which the cover is lowered in the reagent container holder shown in FIG. 20. FIG. [Figure 22] FIG. 11 is a perspective view showing a large-sized reagent container according to a second embodiment of the present invention. [Diagram 23] FIG. 11 is a top view showing a large reagent container according to a second embodiment of the present invention. [Figure 24] FIG. 11 is a vertical sectional view showing a large-sized reagent container according to a second embodiment of the present invention. [Diagram 25] FIG. 11 is a perspective view showing a small-sized reagent container according to a second embodiment of the present invention. [Figure 26] FIG. 11 is a top view showing a small-sized reagent container according to a second embodiment of the present invention. [Figure 27] FIG. 11 is a vertical sectional view showing a small-sized reagent container according to a second embodiment of the present invention. [Figure 28]1A is a diagram showing a state in which a reagent container 200 according to another embodiment of the present invention is installed in a measurement unit 400. FIG. 1B is a diagram showing a state in which an aspirating tube 252 is inserted from above into a reagent container 200 according to another embodiment of the present invention. [Figure 29] 1 is a diagram showing a reagent container 200 according to another embodiment of the present invention, comprising a reagent containing portion 10 and a frame 20. FIG. [Diagram 30] 1 shows that the reagent containing section 10 is formed in a hollow bag shape, and the frame 20 is provided with an opening 21, a mounting member 22, and a movement restricting section 23. FIG. [Diagram 31] 2 is a top view of the movement restricting portion 23. FIG. [Diagram 32] 13 shows that the reagent aspirating section 250 includes a plurality of reagent container holders each capable of holding one reagent container 200. FIG. [Diagram 33] 13 is a diagram showing that the storage section 260 includes a first insertion section 261 into which the reagent storage section 10 of the reagent container 200 is inserted, and a second insertion section 262 into which the movement restricting section 23 of the reagent container 200 is inserted. [Diagram 34] FIG. 2 is a diagram showing a container 260 in which a reagent container 200 is set. [Diagram 35] FIG. 2 is a vertical cross-sectional view of a reagent container holder 250a. [Diagram 36] FIG. 13 is a conceptual diagram illustrating an analysis method according to a fourth embodiment of the present invention. [Figure 37] FIG. 13 is a schematic diagram for explaining waveform data used in the analysis method of the fourth embodiment. [Figure 38] FIG. 13 is a diagram illustrating the conversion into a digital signal by an A / D converter in the analysis method according to the fourth embodiment. [Figure 39] FIG. 13 is a diagram illustrating waveform data obtained by sampling in an analysis method according to a fourth embodiment. [Diagram 40] FIG. 13 is a schematic diagram showing an example of a method for generating training data used to train a deep learning algorithm for determining the type of component in a sample in the analysis method of the fourth embodiment. [Diagram 41]FIG. 13 is a diagram showing an example of label values ​​in the analysis method according to the fourth embodiment. [Diagram 42] FIG. 13 is a schematic diagram showing an example of a method for analyzing waveform data of components in a specimen in an analysis method according to a fourth embodiment. [Diagram 43] 13 is a block diagram showing an example of the configuration of another sample preparation unit different from the sample preparation unit shown in FIG. 12. FIG. [Diagram 44] 1 is a flowchart showing a first operation example of the present analysis method. [Diagram 45] 13 is a flowchart showing a second operation example of the present analysis method. [Figure 46] 13 is a flowchart showing a third operation example of the present analysis method. [Figure 47] FIG. 13 is a block diagram showing a configuration example of a fifth embodiment. [Figure 48] FIG. 2 is a schematic diagram illustrating an example of the configuration of a parallel processing processor. [Figure 49] FIG. 1 is a first diagram showing an overview of arithmetic processing executed in a parallel processing processor based on the control of analysis software running on the processor. [Figure 50] This is the second figure following Figure 49. [Figure 51] This is the third figure following Figure 50. [Figure 52] 1 is a flow chart showing steps for classifying white blood cells into subpopulations using a first fluorescent dye and a second fluorescent dye. [Figure 53] 13 is a flowchart showing the steps of a sixth embodiment. [Figure 54] 13 is a flowchart showing the steps of a seventh embodiment. [Figure 55] 13 is a flowchart showing steps of an eighth embodiment. [Figure 56(a)] 1 shows a side scattering-red fluorescence scattergram when a pre-basophil separation sample in Reference Example 1 was measured. [Figure 56(b)] 1 shows a side scattering-red fluorescence scattergram when a sample after basophil separation in Reference Example 1 is measured. [Fig. 56(c)]1 shows a side scattering-blue-violet fluorescence scattergram when a pre-basophil separation sample was measured in Reference Example 1. [Figure 56(d)] 1 shows a side scattering-blue-violet fluorescence scattergram when a sample after basophil separation in Reference Example 1 is measured. [Figure 57(a)] 1 shows a side scattering-blue-violet fluorescence scattergram when a blood sample taken 4 hours after collection in Reference Example 2 was measured. [Figure 57(b)] 1 shows a side scattering-blue-violet fluorescence scattergram when a blood sample taken 48 hours after collection in Reference Example 2 was measured. [Fig. 57(c)] 1 shows a side scattering-blue-violet fluorescence scattergram when a blood sample taken 72 hours after collection in Reference Example 2 was measured. [Figure 58(a)] FIG. 57(b) is a side scattering-red fluorescence scattergram in which cells gated on the scattergram of FIG. 57(a) in Reference Example 2 are plotted. [Figure 58(b)] FIG. 57(b) is a side scattering-red fluorescence scattergram in which cells gated on the scattergram in FIG. 57(b) in Reference Example 2 are plotted. [Fig. 58(c)] FIG. 57(c) is a side scattering-red fluorescence scattergram in which cells gated on the scattergram in FIG. 57(c) in Reference Example 2 are plotted. [Figure 59(a)] 1 is a side scattering-blue-violet fluorescence scattergram of Example 1. [Figure 59(b)] 1 is a side scatter-red fluorescence scattergram of Example 1. [Figure 60(a)] 1 is a blue-violet fluorescence histogram of Example 2. [Figure 60(b)] 1 is a side scattering-blue-violet fluorescence scattergram of Example 2. [Fig. 60(c)] 1 is a side scatter-red fluorescence scattergram of Example 2. [Figure 61] 1 is a graph showing the change in basophil count over time after blood collection. [Figure 62] 1 shows side scattering-red fluorescence scattergrams in Example 2 and Comparative Example 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present invention will be described in more detail below with reference to the drawings. Note that the following description is illustrative in all respects and should not be construed as limiting the present invention.

[0010] (First embodiment) Fig. 1 is a perspective view showing an analysis system according to a first embodiment of the present invention. As shown in Fig. 1, an analysis system 4000 according to the first embodiment separately includes a measurement device (hereinafter referred to as a measurement unit) 400 and an analysis device (hereinafter referred to as an analysis unit) 300X. In this embodiment, the analysis unit 300X is, for example, a PC (personal computer) in which software for analyzing a sample to be measured is installed.

[0011] The measurement unit 400 is a unit for measuring a specimen, and includes a flow cytometer. In the measurement unit 400, a specimen is mixed with a reagent to prepare a measurement specimen. A reagent containing a plurality of fluorescent dyes corresponding to a plurality of wavelengths is used to prepare the measurement specimen. Each of a plurality of cells in the measurement specimen is stained with a plurality of fluorescent dyes. In the analysis of cells contained in a specimen by this analysis system, cells that can be stained with a plurality of fluorescent dyes are analyzed. That is, the preparation of the measurement specimen in this embodiment is intended to stain one cell with a plurality of fluorescent dyes, and the cells to be measured are, for example, lymphocytes, monocytes, eosinophils, neutrophils, basophils, etc.

[0012] The prepared measurement sample is measured by a flow cytometer. Optical signals corresponding to each of the multiple fluorescent dyes and multiple signals related to forward scattered light and side scattered light are acquired from cells in the measurement sample irradiated with light. Each of the acquired optical signals is A / D converted to acquire digital data. The analysis unit 300X analyzes the digital data acquired by the measurement unit 400. The analysis unit 300X performs at least one of classification and counting of cells in the specimen using multiple digital data corresponding to at least the side scattered light and the multiple wavelengths of fluorescence. The analysis unit 300X also controls the operation of the measurement unit 400.

[0013] Fig. 2 is a schematic diagram showing the configuration of the measurement unit according to the first embodiment of the present invention. As shown in Fig. 1 and Fig. 2, the measurement unit 400 includes a sample preparation section 440 having a chamber 420 and a liquid delivery mechanism 430, a specimen suction mechanism 450, and a flow cytometer detection section (FCM detection section) 460 as a detection section that acquires signals emitted from cells.

[0014] The specimen aspiration unit 450 is a mechanism for aspirating the specimen in the specimen container T, and has a specimen aspiration nozzle 451. The specimen aspiration nozzle 451 can penetrate a specimen container sealed with a lid. The specimen aspiration mechanism 450 can move the specimen aspiration nozzle 451 to insert the specimen aspiration nozzle 451 into the specimen container, and can move in the XY directions to move the specimen aspiration nozzle 451 to a position above the chamber 420. The specimen aspiration mechanism 450 has a metering unit 452 (e.g., a syringe pump) for aspirating and discharging the specimen by the specimen aspiration nozzle 451.

[0015] The liquid sending mechanism 430 includes a liquid sending tube 431 and a liquid sending section 432 for injecting the reagent 12 from the reagent container 200 to the chamber 420 via the liquid sending tube 431. The liquid sending mechanism 430 is a mechanism for sending the reagent from the reagent container 200 to the chamber 420 via the liquid sending tube 431 provided between the reagent container 200 and the chamber 420, which is attached to a reagent container holder 60 (see FIGS. 13 to 15) described later. The reagent container 200 containing the reagent 12 containing both a first fluorescent dye and a second fluorescent dye is attached to the reagent container holder 60. The liquid sending mechanism 430 sends the reagent from the reagent container 200 to the chamber 420. Cells are stained with a first compound constituting the first fluorescent dye and a second compound constituting the second fluorescent dye.

[0016] An aspirating tube 64 constituting one end of a liquid supply tube 431 is inserted into the reagent container 200. The other end of the liquid supply tube 431 is connected to the chamber 420. The aspirating tube 64 may have a sharp tip so that it can penetrate a sealing film (also called a sealing member) of the reagent container 200 attached to the reagent container holder 60.

[0017] The liquid sending unit 432 of the liquid sending mechanism 430 includes a pump 433 as a metering unit that generates a negative pressure for drawing the reagent 12 from the reagent container 200 into the liquid sending tube 431 and a positive pressure for supplying the drawn-in reagent to the chamber 420. The pump 433 can be, for example, a syringe pump or a diaphragm pump. The liquid sending mechanism 430 may include a plurality of valves V1 and V2. For example, when the metering unit 433 configured with a syringe pump or a diaphragm pump draws the reagent from the reagent container, the valve V1 is opened and the valve V2 is closed. The metering tool 433 generates a negative pressure, so that the reagent is filled in the flow path between the valves V1, V2, and the metering unit 433. When the filled reagent is to be supplied to the chamber 420, the valve V1 is closed and the valve V2 is opened, and the metering unit 433 generates a positive pressure. As a result, the reagent in the reagent container 200 is supplied to the chamber 420.

[0018] The chamber 420 is a container in which a reagent and a specimen are mixed to prepare a measurement specimen. In the chamber 420, one reagent 12 containing both the first and second fluorescent dyes is mixed with the specimen to prepare a measurement specimen in which cells are stained with the first and second fluorescent dyes. The measurement unit 400 is provided with one or more chambers 420. The chamber 420 is connected to the waste liquid chamber 36 via a valve 37. After the measurement by the FCM detection unit 460 is completed, the measurement specimen remaining in the chamber 420 is discarded into the waste liquid chamber 36. In addition, the chamber 420 is washed by a washing mechanism (not shown) before the next measurement specimen is prepared, and the liquid after washing is discarded into the waste liquid chamber.

[0019] The measurement unit 400 is provided with one or more reagent container holders 60. Reagent containers 200 that contain a reagent including both a first fluorescent dye that is excited by light and emits fluorescence of a first wavelength, and a second fluorescent dye that is excited by light and emits fluorescence of a second wavelength, are attached to the reagent container holder 60. In the example shown in Fig. 2, a reagent containing the first fluorescent dye and the second fluorescent dye is contained in one reagent container 200.

[0020] The reagent container 200 is a container that contains a reagent. The reagent container 200 has an opening into which a piercer (aspiration tube) 64 connected to a first end of a liquid delivery tube 431 of the liquid delivery mechanism 430 is inserted. Before the reagent container 200 is attached to the reagent container holder 60, the opening is covered with, for example, a sealing film. A piercer 64 is inserted into the opening of the reagent container 200 attached to the reagent container holder 60. The inserted piercer 64 is fixed at a predetermined position in the reagent container 200. The piercer 64 inserted into the reagent container 200 is fixed at the above-mentioned predetermined position, for example, while the reagent container 200 is attached to the reagent container holder 60. Furthermore, the piercer 64 is fixed at the above-mentioned predetermined position at least while measurements of a plurality of different specimens are being performed (i.e., while a plurality of different measurement samples are being prepared).

[0021] The FCM detection unit 460 acquires first and second signals corresponding to the fluorescence of the first and second wavelengths emitted from cells stained with the first and second fluorescent dyes, respectively. The FCM detection unit 460 irradiates light onto the measurement sample flowing through the flow cell. The FCM detection unit 460 may, for example, include a plurality of light sources each corresponding to each wavelength, or may be configured to irradiate light of a single wavelength and detect fluorescence from a plurality of fluorescent dyes excited by the light of the single wavelength. By irradiating the measurement sample with light, optical signals corresponding to the first fluorescent dye and the second fluorescent dye, respectively, are detected. Each optical signal is A / D converted to acquire digital data. The acquired digital data is analyzed by the analysis unit 300X (see FIG. 1).

[0022] In the first embodiment, a liquid delivery pipe 431 is provided between the reagent container 200 and the chamber 420, and a liquid delivery unit 432 delivers the reagent in the reagent container 410 to the chamber 420 via the liquid delivery pipe 431. Therefore, in the first embodiment, the process of aspirating the reagent from the reagent container with a nozzle, moving the nozzle that has aspirated the reagent to the location of the chamber, and discharging the reagent into the chamber (for example, see Non-Patent Document 1) is not necessary.

[0023] The analysis unit 300X (see FIG. 1) performs at least one of classification and counting of cells based on the first and second signals, respectively, corresponding to the fluorescence of the first and second wavelengths emitted from the cells stained with the first and second fluorescent dyes.

[0024] FIG. 3 is a flow chart showing the procedure of the measurement sample preparation process by the analysis system of the first embodiment. The measurement sample preparation process by the analysis system 4000 will be described with reference to FIGS. 1, 2 and 3. In the measurement sample preparation process by the analysis system 4000, first, the measurement unit 400 dispenses a specimen into the chamber 420 (step S1). Next, the reagent is injected into the chamber 420 through the liquid supply pipe 431 connecting the reagent container 410 and the chamber 420 (step S2). Next, in the chamber 420, the specimen and the reagent containing the first and second fluorescent dyes are mixed to prepare a measurement sample (step S3). Next, the measurement sample prepared in the chamber 420 is sent to the FCM detection unit 460 and irradiated with light to obtain optical signals corresponding to the side scattered light and the first and second fluorescent dyes (step S4). Next, the data generated from the obtained optical signals is analyzed by the analysis unit 300X (step S5). Then, the analysis unit 300X provides the analysis results (step S6).

[0025] According to the first embodiment, the first and second fluorescent dyes for reacting the first and second fluorescent dyes with the sample can be supplied to the chamber 420 through the liquid supply tube 431. The liquid supply tube 431 is a dedicated flow path for supplying only the reagent 12 containing the first and second fluorescent dyes to the chamber 420. Since the inside of the liquid supply tube 431 can be constantly kept filled with the reagent 12, the quantification by the quantification unit (pump) 433 can be performed quickly, thereby increasing the processing speed. Furthermore, since the liquid supply tube 431 is a dedicated flow path for supplying the reagent 12, there is no need to prevent contamination between different reagents, and cleaning is not required. The absence of cleaning also contributes to an increase in processing speed.

[0026] The advantages of the first embodiment become even clearer in comparison with, for example, Non-Patent Document 1. For example, the flow cytometer described in Non-Patent Document 1 uses a 96-well plate as a reaction vessel. The 96-well plate is a consumable item, and is removed from the flow cytometer and discarded after use. Therefore, the flow cytometer described in Non-Patent Document 1 is configured such that the reagent vessel and the reaction vessel are connected by a liquid delivery tube, and the process of aspirating and discharging the reagent cannot be eliminated.

[0027] In addition, for the measurement by the flow cytometer described in Non-Patent Document 1, a reagent (Non-Patent Document 2: AQUIOS Tetra-1 Panel) containing four types of fluorescent dyes corresponding to four markers (CD45, CD3, CD4, CD8) in cells in a blood sample is used. Each of these fluorescent dyes is attached to an antibody corresponding to each of the above markers. In other words, the reagent of Non-Patent Document 2 stains cells in a blood sample by antibody-antigen reaction. In the case of such a reagent, the reaction takes time, so it takes time to measure one sample. Specifically, Non-Patent Document 1 describes that it takes about 20 minutes to measure one sample, including preparation of the measurement sample. Since the technology of Non-Patent Document 1 requires a long measurement time per sample (especially the time required for antibody-antigen reaction), the technology of Non-Patent Document 1 shortens the total processing time per sample by preparing measurement samples of multiple samples in parallel in multiple wells of a consumable 96-well plate. In other words, the processing capacity (throughput) per unit time is improved. In contrast, in the first embodiment, a reagent containing first and second fluorescent dyes, the compounds themselves of which stain the cytoplasm, nucleic acid, and DNA of cells, is used as the reagent 12. Such fluorescent dyes have a faster reaction for staining than antibody reagents, and the time required to prepare a measurement sample for one specimen is, for example, less than one minute. Therefore, according to the first embodiment, an analysis device capable of performing measurements using multiple fluorescent labels with high processing power can be provided. Second embodiment

[0028] The analysis system 4000 of the second embodiment is a multi-item automated blood cell analyzer that performs at least one of counting and analyzing cells in a blood sample. FIG. 4 is a block diagram showing the configuration of a measurement unit 400 of the analysis system 4000 (see FIG. 1) of the second embodiment. The measurement unit 400 includes a sample preparation section 440, an apparatus mechanism section 455, a sample suction mechanism 450, an FCM detection section 460, an RBC / PLT detection section 461, an HGB detection section 462, and a measurement unit control section 480. The RBC / PLT detection section 461 is an electrical resistance type detection section that introduces a measurement sample prepared from blood and a diluent into an aperture and counts red blood cells (RBC) and platelets (PLT) by detecting a change in electrical resistance that occurs when cells pass through the aperture. The HGB detection section 462 measures the hemoglobin concentration in blood by the SLS hemoglobin method. The HGB detection unit 462 measures the hemoglobin concentration in blood by irradiating a measurement sample prepared from blood and an SLS hemolyzing agent with light of 555 nm, which is the absorption wavelength of SLS hemoglobin, and measuring the absorbance. Hereinafter, the FCM detection unit 460, the RBC / PLT detection unit 461, and the HGB detection unit 462 may be collectively referred to as "detection units 460 to 462."

[0029] The specimen aspirating mechanism 450 aspirates the specimen from a specimen container and dispenses the aspirated specimen into a chamber of the sample preparation section 440. The sample preparation section 440 includes a chamber for mixing the specimen with a reagent, and a reagent container holder 60 in which a reagent container is placed. The sample preparation section 440 sends the reagent from a reagent container placed in the reagent container holder 60 to the chamber via a liquid sending tube, which will be described later. The specimen and reagent are mixed in the chamber to prepare a measurement sample.

[0030] The device mechanism section 455 includes motors and actuators that move each part of the measurement unit 400. The device mechanism section 455 includes, for example, a mechanism that moves a blood collection tube T (see FIG. 5) described below in the vertical direction.

[0031] The measurement unit control unit 480 includes an analog processing unit 481 that processes the analog signal output from the FCM detection unit 460, an A / D conversion unit 481a that converts the analog signal output from the analog processing unit 481 into a digital signal, an analog processing unit 482 that processes the analog signal output from the RBC / PLT detection unit 461, an A / D conversion unit 482a that converts the analog signal output from the analog processing unit 482 into a digital signal, an analog processing unit 483 that processes the analog signal output from the HGB detection unit 462, an A / D conversion unit 483a that converts the analog signal output from the analog processing unit 483 into a digital signal, and an IF unit (interface unit) 484 electrically connected to each A / D conversion unit 481a, 482a, 483a. Furthermore, the measurement unit control unit 480 includes an interface (IF) section 488 electrically connected to the sample preparation section 440, the device mechanism section 455, the specimen suction mechanism 450, the FCM detection section 460, the RBC / PLT detection section 461 and the HGB detection section 462, a bus 485 electrically connected to each of the IF sections 484, 488, and an IF section 489 electrically connecting the bus 485 to the analysis unit 302X.

[0032] Fig. 5 shows a fluid circuit including a specimen suction mechanism 450, a sample preparation section 440, and detection sections 460 to 462. The sample preparation section 440 shown in Fig. 4 includes a first sample preparation section 440A for preparing a first measurement sample for optical measurement by the FCM detection section 460, and a second sample preparation section 440B for preparing a second measurement sample for electrical resistance-type measurement by the RBC / PLT detection section and a third measurement sample for hemoglobin measurement by the HGB detection section 462 (see Fig. 5).

[0033] The first sample preparation section 440A has a first chamber 420. The first chamber 420 is connected to reagent containers R1 and R2. The reagent container R1 contains a hemolytic agent that shrinks red blood cells. The reagent container 200 contains a white blood cell staining reagent that contains a fluorescent dye. The reagent container R2 contains a diluent. The reagent container R2 will be described later.

[0034] The white blood cell staining reagent contained in the reagent container 200 contains a first fluorescent dye and a second fluorescent dye. The first fluorescent dye and the second fluorescent dye will be described later.

[0035] The second sample preparation section 400B has a second chamber 55. The second chamber 55 is connected to a reagent container R2 and a reagent container R3. The reagent container R2 is provided in common with the first sample preparation section 440A. The reagent container R3 contains an SLS hemolyzing agent for hemolyzing red blood cells and preparing a sample for measurement by the SLS hemoglobin method.

[0036] The reagent container 200 containing the white blood cell staining reagent is held by the reagent container holder 60. The reagent container holder 60 is provided with an aspirating tube 64 for aspirating the nucleic acid staining reagent in the reagent container 200, and an aspirating tube lifting mechanism 65 for raising and lowering the aspirating tube 64. The tip of the aspirating tube 64 can penetrate (puncture) the sealing material of the reagent container 200. A cover 63 is connected to the aspirating tube lifting mechanism 65. When the aspirating tube lifting mechanism 65 is lowered and the aspirating tube 64 penetrates (punctures) the sealing material of the reagent container 200, the cover 63 also descends and covers the reagent container 200. When the aspirating tube lifting mechanism 65 rises, the cover 63 also rises, and the reagent container 200 becomes removable from the outside.

[0037] A liquid delivery mechanism 430 is provided between the suction tube 64 and the first chamber 420. The liquid delivery mechanism 430 includes a liquid delivery tube 431 and a metering block 432. One end of the liquid delivery tube 431 is formed by the suction tube 64, and the other end is connected to the first chamber 420. The metering block 432 includes a metering unit 30 and electromagnetic valves V1 and V2. A syringe pump is used as the metering unit 30. Instead of the syringe pump, for example, a diaphragm pump can also be used. The electromagnetic valves V1 and V2 open and close the flow path. When the leukocyte staining reagent in the reagent container 200 is delivered to the chamber 420, the electromagnetic valve V1 is opened and the electromagnetic valve V2 is closed, and the metering unit 30 applies negative pressure to the liquid delivery tube 431. As a result, the white blood cell staining reagent is sucked into the liquid supply tube 431 from the tip of the suction tube 64, and a fixed amount of the white blood cell staining reagent is filled in the flow path between the electromagnetic valves V1, V2, and the quantification unit 30. Next, with the electromagnetic valve V1 closed and the electromagnetic valve V2 open, the quantification unit 30 applies positive pressure to the liquid supply tube 431. As a result, the fixed amount of the white blood cell staining reagent that has filled the flow path between the electromagnetic valves V1, V2, and the quantification unit 30 is pushed out, and the white blood cell staining reagent is supplied to the chamber 420 through the liquid supply tube 431.

[0038] A metering unit 22 and electromagnetic valves V3 and V4 are provided in the flow path between the reagent container R1 containing the hemolytic agent and the first chamber 420. A syringe pump is used as the metering unit 22. Instead of the syringe pump, for example, a diaphragm pump can be used. The electromagnetic valves V3 and V4 open and close the flow path. The metering unit 22 and the electromagnetic valves V3 and V4 quantitatively send the hemolytic agent in the reagent container R1 to the first chamber 420 in the same manner as the electromagnetic valves V1 and V2 and the metering unit 30 described above.

[0039] A metering unit 33 and electromagnetic valves V5 and V6 are provided in the flow path between the reagent container R2 that contains the diluent and the first chamber 420. A syringe pump is used as the metering unit 33. Instead of the syringe pump, for example, a diaphragm pump can also be used. The electromagnetic valves V5 and V6 open and close the flow path. The metering unit 33 and the electromagnetic valves V5 and V6 quantitatively send the diluent in the reagent container R2 to the first chamber 420.

[0040] A waste liquid chamber 36 that stores unnecessary solution is connected to the first chamber 420. Between the first chamber 420 and the waste liquid chamber 36, an electromagnetic valve V7 that opens and closes the flow path is provided.

[0041] The first chamber 420 is connected to a pump 56A that supplies air into the first chamber 420 to agitate the liquid in the first chamber 420 .

[0042] A metering unit 38 and electromagnetic valves V8 and V9 are provided in the flow path between the reagent container R2 containing the diluent and the second chamber 55. A syringe pump is used as the metering unit 38. Instead of the syringe pump, for example, a diaphragm pump can be used. The electromagnetic valves V8 and V9 open and close the flow path. The metering unit 38 and the electromagnetic valves V8 and V9 quantitatively send the diluent in the reagent container R2 to the second chamber 55. A waste liquid chamber 41 containing unnecessary solution is connected to the second chamber 55. An electromagnetic valve V10 is provided between the second chamber 55 and the waste liquid chamber 41 to switch the flow path between a flow path from the second chamber 55 to the waste liquid chamber 41 and a flow path from the second chamber 55 to the RBC / PLT detection unit 461 and the HGB detection unit 462. The electromagnetic valve V13 will be described later.

[0043] A metering unit 39 and electromagnetic valves V11 and V12 are provided in the flow path between the reagent container R3 containing the SLS hemolytic agent and the second chamber 55. A syringe pump is used as the metering unit 39. Instead of the syringe pump, for example, a diaphragm pump can also be used. The electromagnetic valves V11 and V12 open and close the flow path. The metering unit 39 and the electromagnetic valves V11 and V12 quantitatively send the SLS hemolytic agent in the reagent container R3 to the second chamber 55.

[0044] The second chamber 55 is connected to a pump 56B that supplies air into the second chamber 55 in order to agitate the liquid in the second chamber 55 .

[0045] The specimen suction mechanism 450 has an aspirating tube 20 and a quantification unit 21. The aspirating tube 20 has a sharp tip. When the specimen suction mechanism 450 lowers the blood collection tube 100, the aspirating tube 20 punctures the cover 100a that covers the blood collection tube 100 and is inserted inside. When the aspirating tube 20 is inserted inside the blood collection tube 100, the quantification unit 21 generates negative pressure, so that the blood sample contained in the blood collection tube 100 is aspirated into the aspirating tube 20. The specimen suction mechanism 450 moves the aspirating tube 20 upward to remove the aspirating tube 20 from the blood collection tube 100, and moves the aspirating tube 20 horizontally above the first chamber 420. The specimen suction mechanism 450 lowers the aspirating tube 20 relative to the first chamber 420, and the quantification unit 21 generates positive pressure to discharge the aspirated blood sample into the first chamber 420. The specimen suction mechanism 450 moves the suction tube 20 upward and horizontally above the second chamber 55 , and discharges the blood sample into the second chamber 55 in the same manner as in the first chamber 420 .

[0046] The first chamber 420 is connected to the FCM detection unit 460 (see FIG. 4). The blood sample discharged into the first chamber 420 is mixed with the white blood cell staining reagent contained in the above-mentioned reagent container 200 and the hemolytic agent contained in the reagent container R1 to prepare a measurement sample. More specifically, a measurement sample is prepared in which red blood cells are hemolyzed by the hemolytic agent and white blood cells are stained with a first fluorescent dye and a second fluorescent dye. Such a measurement sample is prepared, for example, as follows. First, a hemolytic agent is supplied to the first chamber 420, and then the blood sample is discharged into the first chamber 420. Air is supplied to the first chamber 420, and the mixture is stirred. As a result, the red blood cells are hemolyzed by the hemolytic agent. Next, a white blood cell staining reagent is supplied to the first chamber 420. Air is supplied to the first chamber 420, and the mixture is stirred. A reaction proceeds in the first chamber 420, and staining with the fluorescent dye is performed. The reaction time is, for example, less than 1 minute, more preferably less than 50 seconds, and more preferably less than 45 seconds. This results in a measurement sample in which white blood cells contained in blood are stained with the first fluorescent dye and the second fluorescent dye. The FCM detection unit 460 is connected to a pump (not shown), and the measurement sample in the first chamber 420 is supplied to the FCM detection unit 460 by driving the pump. The FCM detection unit 460 acquires a plurality of optical signals including the fluorescence corresponding to the first fluorescent dye and the fluorescence corresponding to the second fluorescent dye from the white blood cells.

[0047] The second chamber 55 is connected to the RBC / PLT detection unit 461 and the HGB detection unit 462. The electromagnetic valve V13 switches between sending the measurement sample from the second chamber 55 to the RBC / PLT detection unit 46 and sending the measurement sample to the HGB detection unit 462. The RBC / PLT detection unit 461 and the HGB detection unit 462 are connected to a pump (not shown), and the measurement sample in the second chamber 55 is supplied to the RBC / PLT detection unit 461 and the HGB detection unit 462 by driving the pump. The second chamber 55 is used to prepare both the measurement sample for RBC / PLT detection and the measurement sample for HGB detection. An example of the preparation procedure of such a sample will be described. First, a diluent is supplied from the reagent container R2 to the second chamber 55. Next, a blood sample is discharged into the second chamber 55. This results in a measurement sample in which the blood is diluted. This becomes the measurement sample for RBC / PLT detection. A portion of the measurement sample in second chamber 55 is sent to RBC / PLT detection section 461, where electrical resistance detection is performed. Next, an SLS hemolyzing agent is supplied from reagent container R3 to the measurement sample remaining in second chamber 55. This results in a measurement sample in which red blood cells are hemolyzed and hemoglobin is converted to SLS hemoglobin. This measurement sample is sent to HGB detection section 462. Note that in the example of FIG. 5, the measurement sample for RBC / PLT detection and the measurement sample for HGB detection are prepared in the common second chamber 55, but they may be prepared in separate chambers.

[0048] The analysis system 4000 (see FIG. 1) having such a configuration may be configured to be capable of measuring a complete blood count (CBC) consisting of at least eight parameters, namely, red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hemoglobin concentration (HGB), hematocrit value (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). Furthermore, the analysis system 4000 may be configured to be capable of measuring a DIFF item that classifies white blood cells into a plurality of subpopulations in addition to the CBC item.

[0049] <Modifications regarding the sample preparation section> FIG. 6 is a schematic diagram showing another example of the first sample preparation section 440A. In FIG. 6, elements similar to those in FIG. 5 are omitted. In the example shown in FIG. 5, the first sample preparation section 440A is configured to accommodate a reagent containing a first fluorescent dye and a second fluorescent dye in one reagent container 200, and to send the reagent in one reagent container 200 into the chamber 420 by one liquid sending mechanism 430. In the first sample preparation section 440A of the modified example shown in FIG. 6, the reagent containing a first fluorescent dye and the reagent containing a second fluorescent dye are accommodated in individual reagent containers 200A and 200B, respectively, and the reagent in each reagent container 200A and 200B is sent into the chamber 420 by individual first and second liquid sending mechanisms 430a and 430b, respectively.

[0050] 6 includes one or more chambers 420 for mixing two reagents containing either a first or second fluorescent dye attached to a reagent container holder with a specimen to prepare a measurement sample in which cells are stained with the first and second fluorescent dyes. A first reagent container 200A containing a reagent containing the first fluorescent dye is attached to a first holder portion of the reagent container holder 442, and a second reagent container 200B containing a reagent containing the second fluorescent dye is attached to a second holder portion of the reagent container holder 442.

[0051] The first liquid delivery mechanism 430a is provided to deliver the reagent in the first reagent container 200A to the chamber 54. The configuration of the first liquid delivery mechanism 430a is as described with reference to FIG. 3. The second liquid delivery mechanism 430b is provided to deliver the reagent in the second reagent container 200B to the chamber 54. The configuration of the second liquid delivery mechanism 430b is similar to that of the first liquid delivery mechanism. The liquid delivery tubes 431 of the two liquid delivery mechanisms 430a and 430b join in the middle of the flow path and are connected to the chamber 420. Note that, although the example of FIG. 6 shows an example in which two liquid delivery tubes join, the two liquid delivery tubes may be connected to the chamber 420 individually.

[0052] 6, a reagent containing a first fluorescent dye and a reagent containing a second fluorescent dye are contained in different reagent containers 200A and 200B, respectively, and are supplied to chamber 420 by two different liquid delivery mechanisms and mixed with the specimen. With this configuration as well, a system can be constructed in which blood cell components contained in a specimen are stained with two fluorescent dyes.

[0053] Fig. 7 is a configuration explanatory diagram showing an example of the optical system of the FCM detection unit 460. As shown in Fig. 7, the FCM detection unit 460 includes a first light source 4111a and a second light source 4111b having mutually different wavelengths, a flow cell 4113, dichroic mirrors 4118a, 4118b, and 4118c, side scattered light receiving elements 4121a and 4121b, a forward scattered light receiving element 4116, and side fluorescent light receiving elements 4122a and 4122b. In the example of Fig. 7, the light source is composed of a first light source 4111a for exciting a first fluorescent dye and a second light source 4111b for exciting a second fluorescent dye.

[0054] The FCM detection unit 460 acquires the first signal composed of a plurality of signals corresponding to the side scattered light and side fluorescent light by the first light source 4111a, and the second signal composed of a plurality of signals corresponding to the forward scattered light, side scattered light, and side fluorescent light by the second light source 4111b. As described above, the FCM detection unit 460 is connected to the first chamber 420 of the first sample preparation unit 440A (see FIG. 5). The measurement sample prepared in the first chamber 420 is made to flow through the flow cell (sheath flow cell) 4113 of the FCM detection unit 460. In the example of FIG. 7, the measurement sample is made to flow in a direction perpendicular to the paper surface. With the measurement sample flowing through the flow cell 4113, the first light source 4111a and the second light source 4111b irradiate light onto the flow cell 4113. More specifically, light emitted from a first light source 4111a is reflected by a dichroic mirror 4118a and irradiated onto the flow cell 4113. Light emitted from a second light source 4111b is transmitted through a dichroic mirror 4118a and irradiated onto the flow cell 4113.

[0055] The first light source 4111a and the second light source 4111b of the FCM detection unit 460 are not particularly limited, and a light source having a wavelength suitable for exciting a fluorescent dye is selected. The first light source 4111a can irradiate light of a first wavelength, for example, and the second light source 4111b can irradiate light of a second wavelength that is longer than the first wavelength. For example, the first wavelength is 315 to 490 nm, preferably 400 to 450 nm, and more preferably 400 to 410 nm. The second wavelength is 610 to 750 nm, preferably 620 to 700 nm, and more preferably 633 to 643 nm. As such first and second light sources 4111a and 4111b, for example, a semiconductor laser light source, an argon laser light source, a gas laser light source such as a helium-neon laser, a mercury arc lamp, or the like can be used. In particular, a semiconductor laser light source is preferable because it is less expensive than a gas laser light source. As described above, by selecting the first light source 4111a and the second light source 4111b that emit light with wavelength bands that are far apart, it becomes easier to select a dye with a small overlap in the wavelength bands of the fluorescence from the fluorescent dyes corresponding to each light source. If the overlap in the wavelength bands of the fluorescence emitted from the fluorescent dyes is large, it is necessary to deal with the spillover described below, but by selecting a fluorescent dye with a small overlap (or no overlap) in the wavelength bands of the fluorescence, it becomes unnecessary to deal with the spillover.

[0056] The forward scattered light receiving element 4116 is disposed so as to receive forward scattered light emitted from the cells based on the light irradiated from the second light source 4111b. The forward scattered light receiving element 4116 is configured to receive, for example, only forward scattered light corresponding to the light irradiated from the second light source 4111b. The forward scattered light receiving element 4116 is, for example, a photodiode. In this example, the light receiving element 4116 receives forward scattered light of a second wavelength irradiated from the second light source 4111b, but alternatively, it may receive forward scattered light of a first wavelength irradiated from the first light source 4111a.

[0057] The side scattered light receiving element 4121a is disposed so as to receive side scattered light corresponding to the light from the first light source 4111a. The side scattered light (first side scattered light) corresponding to the light emitted from the first light source 4111a is reflected by the dichroic mirror 4118b and received by the side scattered light receiving element 4121a. The side scattered light receiving element 4121a is, for example, a photodiode.

[0058] The side scattered light receiving element 4121b is disposed so as to receive side scattered light corresponding to the light from the second light source 4111b. The side scattered light (second side scattered light) corresponding to the light emitted from the second light source 4111b is reflected by the dichroic mirror 4118c and received by the side scattered light receiving element 4121b. The side scattered light receiving element 4121b is, for example, a photodiode.

[0059] The side fluorescent light receiving element 4122a is disposed so as to receive fluorescence corresponding to the light from the first light source 4111a, i.e., light generated when the first fluorescent dye is excited by the light of the first wavelength from the first light source 4111a. The side fluorescent light (first side fluorescent light) corresponding to the light irradiated from the first light source 4111a passes through the dichroic mirror 4118b and is received by the side fluorescent light receiving element 4122a. The side fluorescent light receiving element 4122a is, for example, an avalanche photodiode.

[0060] The side fluorescent light receiving element 4122b is disposed to receive fluorescence corresponding to the light from the second light source 4111b, that is, light generated by exciting the second fluorescent dye with the light of the second wavelength from the second light source 4111b. The side fluorescent light (second side fluorescent light) corresponding to the light irradiated from the second light source 4111b passes through the dichroic mirror 4118c and is received by the side fluorescent light receiving element 4122b. The side fluorescent light receiving element 4122b is, for example, an avalanche photodiode. Note that a photomultiplier tube may be used as the forward scattered light receiving element 4116, the side scattered light receiving elements 4121a, 4121b, and the side fluorescent light receiving elements 4122a, 4122b.

[0061] The forward scattered light receiving element 4116, the side scattered light receiving elements 4121a, 4121b, and the side fluorescent light receiving elements 4122a, 4122b each output a waveform-shaped electrical signal (also called a received light signal) including a pulse according to the received light intensity. Typically, each pulse of the received light signal corresponds to one cell (e.g., a white blood cell). The output received light signals are each input to an analog processing unit 481. The analog processing unit 481 performs processing such as noise removal and smoothing on the analog signal input from the FCM detection unit 460, and outputs the processed analog signal to an A / D conversion unit 481a. The other analog processing units 482, 483 (see FIG. 4) are configured in the same manner as the analog processing unit 481.

[0062] The A / D conversion unit 481a converts the analog signal output from the analog processing unit 481 into a digital signal. The A / D conversion unit 481a converts the analog signal from the start of sample measurement to the end of measurement into a digital signal. When multiple types of analog signals (for example, analog signals corresponding to forward scattered light intensity, first side scattered light intensity, first fluorescent light intensity, second side scattered light intensity, and second fluorescent light intensity, respectively) are generated by measurement in a certain measurement channel, the A / D conversion unit 481a converts each analog signal from the start of measurement to the end of measurement into a digital signal. For example, five types of analog signals (i.e., a forward scattered light signal, a first side scattered light signal, a first fluorescent light signal, a second side scattered light signal, and a second fluorescent light signal) are input to the A / D conversion unit 481a. The A / D conversion unit 481a converts each of the input analog signals into a digital signal.

[0063] The A / D conversion unit 481a samples analog signals at a predetermined sampling rate (for example, sampling of 1024 points at 10 nanosecond intervals, sampling of 128 points at 80 nanosecond intervals, or sampling of 64 points at 160 nanosecond intervals). The A / D conversion unit 481a performs sampling processing on five types of analog signals corresponding to individual cells flowing through the flow cell 4113, for example, to generate waveform data of a forward scattered light signal, waveform data of a first side scattered light signal, waveform data of a first fluorescent light signal, waveform data of a second side scattered light signal, and waveform data of a second fluorescent light signal for each formed component. The A / D conversion unit 481a assigns an index to each of the generated waveform data. The generated waveform data becomes, for example, a digital signal having signals corresponding to N cells contained in one specimen. As a result, five digital signals corresponding to five types of analog signals (forward scattered light signal, first and second side scattered light signals, and first and second fluorescent light signals) obtained from N cells are generated (see FIG. 39).

[0064] The A / D conversion unit 481a generates waveform data from the analog signal, and also calculates characteristic parameters representing the morphological characteristics of individual cells from the pulse of the analog signal, such as a peak value, a pulse width, and a pulse area.

[0065] Fig. 8 is a configuration explanatory diagram showing another example of the optical system of the FCM detection unit. As shown in Fig. 8, the light source of the FCM detection unit 460x may be composed of one light source 4111 that excites a first fluorescent dye and a second fluorescent dye. In this case, the FCM detection unit 460x acquires the first signal composed of a plurality of signals respectively corresponding to the forward scattered light, the side scattered light, and the side fluorescent light from the light source 4111, and a second signal corresponding to the side fluorescent light from the light source 4111.

[0066] In this case, light of a predetermined wavelength, for example, is irradiated from one light source 4111 onto the flow cell 4113. As in the above example, cells contained in the measurement sample flowing through the flow cell 4113 are stained with multiple types of fluorescent dyes that emit different wavelengths of fluorescence when excited. In this example, multiple types of fluorescent dyes that are excited with different wavelengths when irradiated with light of a predetermined wavelength (for example, 488 nm) are used as the multiple types of fluorescent dyes. Multiple types of fluorescent dyes that are excited with different wavelengths mean, for example, that each fluorescent dye is excited by being irradiated with light from one light source and emits different colors of fluorescence.

[0067] In the optical system of the FCM detection unit 460x shown in Fig. 8, cells contained in a measurement sample are stained with multiple fluorescent dyes, and the cells are irradiated with light from one light source 4111, and each fluorescence emitted from the multiple fluorescent dyes can be detected. In this case, since the light irradiated from one light source 4111 has a single wavelength, when the light is irradiated from one light source 4111 to a cell, one side scattered light corresponding to the single wavelength of the light is emitted. Therefore, in the optical system of the FCM detection unit 460x shown in Fig. 8, one dichroic mirror 4118 is provided as a dichroic mirror, and the other configurations are generally similar to those of the optical system shown in Fig. 7.

[0068] The fluorescence of each fluorescent dye has its own spectrum. Therefore, when the fluorescence is separated into a single wavelength range by an optical system (for example, the dichroic mirror 4118 and the side fluorescent light receiving elements 4122a and 4122b in FIG. 7), the fluorescence emitted from the fluorescent dye other than the one fluorescent dye bound to the target molecule in one cell may leak into each of the side fluorescent light receiving elements 4122a and 4122b. For example, as in the example shown in FIG. 9, when multiple fluorescent dyes are excited by irradiating them with light of a single wavelength, the overlap of the wavelength bands of the fluorescence emitted by each fluorescent dye upon excitation may become large. When multiple fluorescent dyes with overlapping wavelength bands of fluorescence are selected as in FIG. 9 showing an example of overlapping wavelength bands of fluorescence, for example, a measure against the leakage of fluorescence is implemented.

[0069] FIG. 9 is a diagram showing an example in which fluorescence emitted from two different fluorescent dyes leaks into each other. In the example shown in FIG. 9, fluorescence emitted from two dyes, fluorescent dye F1 and fluorescent dye F2, leaks into each other. The vertical axis of the graph shown in FIG. 9 indicates the fluorescence intensity, and the horizontal axis indicates the wavelength. The light emitted from the fluorescent dye F1 is detected by one of the side fluorescent light receiving elements in the wavelength band (a) of FIG. 9. Moreover, the light emitted from the fluorescent dye F2 is detected by the other side fluorescent light receiving element in the wavelength band (b) of FIG. 9. The wavelength band (a) detected by one of the side fluorescent light receiving elements corresponding to the fluorescent dye F1 includes the wavelength band of the fluorescence emitted by the fluorescent dye F2. Moreover, the wavelength band (b) detected by the other side fluorescent light receiving element corresponding to the fluorescent dye F2 includes the wavelength band of the fluorescence emitted by the fluorescent dye F1. Therefore, by performing a correction to remove the portion of the fluorescence in wavelength band (a) (corresponding to fluorescent dye F1) that corresponds to fluorescent dye F2, a more accurate measurement value of the fluorescence from fluorescent dye F1 can be obtained. Also, by performing a correction to remove the portion of the fluorescence in wavelength band (b) (corresponding to fluorescent dye F2) that corresponds to fluorescent dye F1, a more accurate measurement value of the fluorescence from fluorescent dye F2 can be obtained.

[0070] To correct for the fluorescence spillover shown in Fig. 9, a positive control for fluorescent dye F1 and a positive control for fluorescent dye F2 are used. For the positive control for fluorescent dye F1, for example, a control reagent containing particles to which fluorescent dye F1 has been added is used. For the positive control for fluorescent dye F2, for example, a control reagent containing particles to which fluorescent dye F2 has been added is used.

[0071] Measurement of the positive control of fluorescent dye F1 by the analysis system having the optical system illustrated in Fig. 8 is performed as follows before measurement of a specimen. Light (light of a single wavelength) from a light source 4111 is irradiated onto particles in the positive control flowing through a flow cell 4113. Fluorescence generated by irradiating the particles with light is detected by each of the side fluorescent light receiving elements 4122a, 4122b. For example, the side fluorescent light receiving element 4122a corresponds to detection of fluorescence emitted from the fluorescent dye F1, and the side fluorescent light receiving element 4122b corresponds to detection of fluorescence generated from the fluorescent dye F2. In this case, of the fluorescence emitted from the particles in the positive control of fluorescent dye F1, the fluorescence detected by the side fluorescent light receiving element 4122b becomes a spillover. For example, if the fluorescence intensity detected by the side fluorescent light receiving element 4122a among the fluorescence emitted from the particles in the positive control of the fluorescent dye F1 is "100" and the fluorescence intensity detected by the side fluorescent light receiving element 4122b is "5", 5% of the fluorescence from the fluorescent dye F1 leaks into 4122b. For example, if the fluorescence intensity detected by the side fluorescent light receiving element 4122a among the fluorescence emitted from the particles in the positive control of the fluorescent dye F2 is "10" and the fluorescence intensity detected by the side fluorescent light receiving element 4122b is "100", 10% of the fluorescence from the fluorescent dye F2 leaks into the side fluorescent light receiving element 4122a. The results are summarized in Table 1 below. This Table 1 may be hereinafter referred to as the "compensation matrix".

[0072] [Table 1]

[0073] Next, an example of correcting the measured values ​​obtained from a measurement sample containing cells stained with each of the fluorescent dyes F1 and F2 using the compensation matrix will be described. An example of the correction of the measured values ​​is as follows. Corrected fluorescence intensity of fluorescent dye F1 measured by the side fluorescence receiving element 4122a = fluorescence intensity measured by the side fluorescence receiving element 4122a - (5% of the fluorescence intensity when the positive control of fluorescent dye F2 is measured by the side fluorescence receiving element 4122b) Corrected fluorescence intensity of fluorescent dye F2 measured by the side fluorescence receiving element 4122b = fluorescence intensity measured by the side fluorescence receiving element 4122b - (10% of the fluorescence intensity measured by the side fluorescence receiving element 4122a for the positive control of fluorescent dye F1)

[0074] In the above example, the fluorescence intensity when the fluorescence of the positive control for fluorescent dye F1 is detected by the side fluorescent light receiving element 4122a is "100," and the fluorescence intensity when the fluorescence of the positive control for fluorescent dye F2 is detected by the side fluorescent light receiving element 4122b is "100." Therefore, the corrected fluorescence intensities are as follows, respectively. Corrected fluorescence intensity of the fluorescent dye F1 by the side fluorescence receiving element 4122a = Fluorescence intensity measured by the side fluorescence receiving element 4122a 100 - 10 Corrected fluorescence intensity of fluorescent dye F2 by the side fluorescence receiving element 4122b = Fluorescence intensity measured by the side fluorescence receiving element 4122b 100 - 5

[0075] 7 and 8, the optical system detects the fluorescence (side fluorescence) generated to the side of the traveling direction of the irradiated light (approximately 90° to the traveling direction in the examples of Figs. 7 and 8) as the fluorescence generated by exciting the first fluorescent dye and the second fluorescent dye, but is not limited to this example. For example, the fluorescence generated forward of the traveling direction of the irradiated light may be detected, or the fluorescence generated at a certain angle, for example, 20 to 90°, other than a right angle to the traveling direction of the irradiated light may be detected.

[0076] FIG. 10 is a block diagram showing an example of the configuration of the analysis unit 300X. As shown in FIG. 10, the analysis unit 300X is electrically connected to the measurement unit 400 via an interface unit 3006. The interface unit 3006 is, for example, a USB interface. The analysis unit 300X includes a processor 3001, a main memory 3017, a bus 3003, a storage unit 3004, the interface unit 3006, a display unit 3015, and an operation unit 3016. The analysis unit 300X is, for example, configured by a personal computer (see the analysis unit 300X in FIG. 1), and controls the measurement unit 400 of the analysis system 4000 by executing a program stored in the storage unit 3004. The analysis unit 300X executes, for example, a program for analysis, and analyzes data acquired by the measurement unit 400. The analysis unit 300X displays the analysis results on the display unit 3015. In the example described with reference to FIG. 7, for example, the analysis unit 300X may classify cells by inputting waveform data corresponding to at least one, preferably a plurality, of the forward scattered light intensity, the first side scattered light intensity, the first fluorescent light intensity, the second side scattered light intensity, and the second fluorescent light intensity acquired from each cell by the measurement unit 400 using the FCM detection unit 460 into a trained AI algorithm. Alternatively, the analysis unit 300X may classify cells based on characteristic parameters (peak value, pulse width, pulse area) of each cell based on the forward scattered light intensity, the first side scattered light intensity, the first fluorescent light intensity, the second side scattered light intensity, and the second fluorescent light intensity. For example, as a method of classifying particles into multiple types using multiple characteristic parameters, a method can be adopted in which particles are plotted in a multidimensional coordinate space with multiple parameters as axes, at least some particles are classified into multiple groups corresponding to multiple types, the degree of belonging of each particle to each group is calculated based on the distance between the center of gravity of each group and the particle, and the particles are reclassified based on the degree of belonging to the particles, thereby classifying the particles into multiple types. Such classification methods are described, for example, in U.S. Patent No. 5,555,198, which is hereby incorporated by reference.Alternatively, some cells contained in a measurement sample may be classified based on an AI algorithm, while other cells may be classified based on characteristic parameters.

[0077] The processor 3001 is a CPU (Central Processing Unit) and executes a program loaded from the storage unit 3004 to the main memory 3017. The storage unit 3004 is, for example, a hard disk or a solid state drive (SSD). The storage unit 3004 stores, for example, a program for controlling the measurement unit 400, a program for analyzing data acquired by the measurement unit 400, and the like. The display unit 3015 includes a computer screen. The display unit 3015 is electrically connected to the processor 3001 via the interface unit 3006 and the bus 3003. The display unit 3015 displays, for example, the results of an analysis of the data acquired by the measurement unit 400.

[0078] The operation unit 3016 includes a pointing device including a keyboard, a mouse, or a touch panel. A user such as a doctor or a laboratory technician can input a measurement order to the analysis system 4000 and input a measurement instruction according to the measurement order by operating the operation unit 3016. The operation unit 3016 can also receive an instruction from a user to display the test results. A user can operate the operation unit 3016 to view various information related to the test results, such as graphs, charts, and flag information assigned to the sample. The above-mentioned measurement unit 400 is electrically connected to the analysis unit 300X via the interface unit 3006.

[0079] Fig. 11 is a perspective view showing another configuration example of the analysis system 4000. In the configuration examples shown in Figs. 1 to 10, the analysis system 4000 in which the measurement unit 400 and the analysis unit 300X are provided as separate entities is exemplified, but as shown in Fig. 11, the analysis system 4001 may be one in which the analysis section 300X is provided within the measurement unit 400. With this configuration, the entire analysis system 4001 becomes compact.

[0080] Fig. 12 is a block diagram showing the configuration of another example of the measurement unit. In the examples of Fig. 2, Fig. 5, and Fig. 6, an example is shown in which there is one measurement system (also called measurement channel) that can be measured by the FCM detection unit 460. In the configuration example of Fig. 12, an example is shown in which five measurement channels are provided.

[0081] The first sample preparation section 440A includes five chambers 54a to 54e. The chambers 54a to 54e are used in the measurement channels of DIFF, RET, WPC, PLT-F, and WNR, respectively. A hemolytic agent container containing a hemolytic agent and a staining solution container containing a staining solution, which are reagents corresponding to each measurement channel, can be delivered to each chamber 54a to 54e through a flow path. One measurement channel is composed of one chamber and a hemolytic agent container and a staining solution container provided so as to be able to deliver liquid to the chamber. For example, the DIFF measurement channel is composed of a hemolytic agent container containing a DIFF hemolytic agent, which is a DIFF measurement reagent, a staining solution container containing a DIFF staining solution, and a DIFF chamber 54a connected to them through a flow path. The other measurement channels are similarly configured. Here, a case is illustrated in which one measurement channel is configured to supply both one type of hemolytic agent and one type of staining solution to the chamber, but the present invention is not limited to this configuration. For example, the measurement channels may be configured to supply both one type of hemolytic agent and one type of staining solution to different chambers. In other words, one reagent (one type of hemolytic agent and one type of staining solution) may be shared by multiple measurement channels, and for example, two of the five measurement channels may be DIFF measurement channels. Furthermore, the measurement channel may be one, or, for example, a configuration may be provided in which only two DIFF measurement channels are provided.

[0082] The sample suction mechanism 450 (see Figs. 2, 4, and 5) aspirates the blood sample from a sample container T containing the blood sample, moves the device mechanism 455 (see Fig. 4) to an upper position of one of the chambers 54a to 54e corresponding to the measurement channel corresponding to the order, and discharges the aspirated blood sample into the chamber. The sample preparation unit 440 supplies the corresponding hemolytic agent and staining solution to the chamber into which the blood sample is discharged, and prepares a measurement sample by mixing the blood sample, hemolytic agent, and staining solution in the chamber. The prepared measurement sample is supplied from the chamber via a flow path to the FCM detection unit 460, where cells are measured by flow cytometry.

[0083] The above-mentioned measurement channels (DIFF, RET, WPC, PLT-F, WNR) correspond to the measurement items included in the measurement order. For example, DIFF corresponds to the measurement item related to the classification of white blood cells. RET corresponds to the measurement item related to reticulocytes. WPC corresponds to the measurement item related to the measurement of abnormal white blood cells. PLT-F corresponds to the measurement item related to the optical measurement of platelets. WNR corresponds to the measurement item related to white blood cells and nucleated red blood cells. The above-mentioned measurement channels (DIFF, RET, WPC, PLT-F, WNR) are measured by the FCM detection unit 460.

[0084] FIG. 13 is a diagram showing a state in which the cover 442 of the measuring unit 40 is opened. FIG. 14 is a perspective view showing the reagent container holder 60 of the measuring unit 400, and FIG. 15 is a front view showing the reagent container holder 60 shown in FIG. 14. An openable front cover 442 (see FIG. 13) is provided on the front side of the measuring unit 400 (see FIG. 1) of the analysis system 4000. The reagent container holder 60 is disposed on the upper front part of the measuring unit 400, and as shown in FIG. 13, by opening the cover 442, the reagent container holder 60 is exposed and can be accessed by the user. In other words, the user can place the reagent container 200 in the reagent container holder 60 and remove the reagent container 200 from the reagent container holder 60.

[0085] As shown in FIG. 14 and FIG. 15, the reagent container holder 60 includes five holder parts 60a, 60b, 60c, 60d, and 60e, and is configured to hold a total of five (five types) reagent containers 200 (or 300). The reagent containers 200 (or 300) held by the reagent container holder 60 contain different types of reagents (staining solutions) for measuring a plurality of measurement items by the FCM detection unit 460. The color of the reagent container is, for example, black. Depending on the type of reagent, a large (about 100 mL) reagent container 200 (see FIG. 22) and a small (about 20 mL) reagent container 300 (see FIG. 25) are used as the reagent container. The capacity of the reagent container is not limited to the above, and may be, for example, less than 20 ml, less than 100 ml, or more than 100 ml. For example, it may be about 10 ml, about 40 ml, about 80 ml, or about 300 ml. Each of the holder parts 60a-60e is configured to be able to hold, for example, either the reagent container 200 or 300. Thus, the five holder parts 60a-60e each have the same configuration, and for example, large reagent containers 200 can be set in the three holder parts 60a-60c, and small reagent containers 300 can be set in the two holder parts 60d and 60e. Each of the holder parts 60a-60e includes a chassis 61, a reagent container holding part 62, a cover 63 for opening and closing the reagent container holding part 62, the above-mentioned aspirating tube 64, and an aspirating tube lifting mechanism 65.

[0086] The reagent container holder 62 is provided at the lower part of the chassis 61 (see Figs. 14 and 15). As shown in Fig. 15, the reagent container holder 62 has a height H, and as shown in Fig. 16, includes a first receiving part 621 having a width W11, an intermediate receiving part 622 that is continuous with the first receiving part 621 and expands from the first receiving part 621 at a predetermined angle θ2, and a second receiving part 623 that is continuous with the intermediate receiving part 622. As shown in Figs. 16 and 17, the first receiving part 621 can receive a first storage part 210 (310) of the reagent container 200 (300) described later, and has a width (width W11) that prohibits the entry of a user's finger. The finger means, for example, an adult finger having an average thickness, and the width W11 is, for example, 10 mm. The second receiving part 623 has a width W12 that is larger than the width W11. 16 and 17, the first receiving section 621 is disposed at the innermost side (the side in the direction of the arrow Y2) of the reagent container holding section 62. The reagent container 200 (300) is inserted from an entrance section 212 (312) (described later) of the first accommodation section 210 (310) toward the innermost side of the reagent container holding section 62. Therefore, the reagent container holding section 62 is configured to hold the reagent container 200 (300) in a state where the entrance section 212 (312) of the reagent container 200 (300) is inserted so that the entrance section 212 (312) is at the innermost side (the side in the direction of the arrow Y2).

[0087] 16 to 18, the reagent container holder 62 includes a pair of guide members 627 that guide both side surfaces 214 (314) of the first storage portion 210 (310) of the reagent container 200 (300) to the first receiving portion 621. The guide members 627 include a first guide portion 627a that guides the first storage portion 210 (310) of the reagent container 200 (300) to the first receiving portion 621, an intermediate guide portion 627b that corresponds to the intermediate receiving portion 622, and a second guide portion 627c that guides a second storage portion 220 (320) (to be described later) of the reagent container 200 (300) to the second receiving portion 623. The guide members 627 are formed by a part (both inner side surfaces) of the chassis 61. The first receiving portion 621, the intermediate receiving portion 622, and the second receiving portion 623 are each formed by a space between a pair of corresponding first guide portions 627a, a space between a pair of intermediate guide portions 627b, and a space between a pair of second guide portions 627c. Therefore, the width W11 of the first receiving portion 621 is equal to the width between the pair of first guide portions 627a, and the width W12 of the second receiving portion 623 is equal to the width between the pair of second guide portions 627c.

[0088] The pair of guide members 627 have a height H (see FIG. 14) that is approximately equal to a height H1 (see FIGS. 24 and 27) of both side surfaces 214 (314) of the first storage section 210 (310) of the reagent container 200 (300), and are configured to be able to guide both side surfaces 214 (314) of the first storage section 210 (310) of the reagent container 200 (300) from the lower end to the upper end. Furthermore, the pair of guide members 627 have a shape that reflects the outer shape of the first storage section 210 (310), and are configured to be able to guide the entire both side surfaces 214 (314) of the first storage section 210 (310) of the reagent container 200 (300). Moreover, the intermediate guide portion 627b and the second guide portion 627c have a shape that reflects the outer shape of the large reagent container 200, and are configured to be able to guide both side surfaces of the tip side (first storage portion 210 side) half of the second storage portion 220. The second storage portion 220 of this large reagent container 200 has a width W22 that is larger than the width W21 of the first storage portion 210, and the first guide portion 627a is provided with a width W11 that is smaller than the width W22 of the second storage portion 220.

[0089] 18, the reagent container holding section 62 includes a support section 624 that supports the reagent container 200 (300) and a rotation mechanism 625 that rotatably supports the support section 624. The support section 624 is a plate-like member integrally having a front side section 624a that comes into contact with the front surface of the reagent container 200 (300) (the tip surface of the first storage section 210 (310), see FIGS. 16 and 17) and a lower side section 624b that comes into contact with the lower surface of the reagent container 200 (300). That is, the support section 624 is formed to have a shape corresponding to the shape of the reagent container 200 (300). The rotation mechanism 625 is configured such that a protrusion 624c provided on the support portion 624 is inserted into an annular bearing 625a provided on the inner surface of the chassis 61, thereby enabling the support portion 624 to rotate around the position of the protrusion 624c (bearing 625a) as the rotation center.

[0090] In addition, a locking portion 626 is provided inside the chassis 61 to lock the rotating support portion 624 by abutting against a front portion 624a of the support portion 624. A magnet is provided in the locking portion 626, and the locking portion 626 is configured to hold the front portion 624a (support portion 624) when in contact with the front portion 624a of the support portion 624. As a result, the support portion 624 is configured to move between a placement position P1 (see FIG. 19) where the lower portion 624b is horizontal (the lower surface of the reagent container 200 (300) is horizontal) and a set position Q1 (see FIG. 20) where the front portion 624a is vertical. As shown in FIG. 20, when placed at this set position Q1, an entrance portion 212 (312) (see FIGS. 16 and 17) of the reagent container 200 (300) described later is configured to be horizontal (perpendicular to the suction tube 64).

[0091] 19, the cover 63 is disposed so as to protrude from each of the holder portions 60a to 60e (chassis 61) toward the front (the direction of arrow Y1), and is attached to an aspirating tube lifting mechanism 65. The aspirating tube lifting mechanism 65 enables the cover 63 to move between an elevated position P2 (see FIG. 19) that opens the reagent container holding portion 62, and a lowered position Q2 (see FIG. 21) that covers (closes) the reagent container holding portion 62. Thus, the cover 63 is configured to permit insertion and removal of the reagent container 200 (300) in the elevated position P2, and to prohibit insertion and removal of the reagent container 200 (300) in the lowered position Q2.

[0092] 15, a window 631 consisting of an opening is provided at a predetermined position of the cover 63. As shown in FIG. 21, when the cover 63 is in the lowered position Q2 where the reagent container holder 62 is covered (closed), the user can view the label 250 (350, see FIG. 25) attached to the reagent container 200 (300) through the window 631. A mark for identifying the type (type of reagent) of the reagent container 200 (300) is printed on the label 250 (350) at a position visible through the window 631. A label 632 on which a mark for identifying the type (type of reagent) of the reagent container 200 (300) to be set in the reagent container holder 62 is printed is attached to the cover 63. That is, reagent containers 200 (300) containing predetermined types of reagents are set in the five holder parts 60a to 60e, and correspondingly, labels 632 identifying the types of reagents to be set are affixed to the covers 63 of the holder parts 60a to 60e. With this, when the reagent containers 200 (300) are set in the reagent container holding part 62 (when the cover 63 is lowered to the lowered position Q2), it is possible to check from the labels 632 affixed to the covers 63 and the labels 250 (350) visible through the windows 631 whether the correct reagents have been set in the holder parts 60a to 60e.

[0093] As shown in Figures 16 and 20, the aspirating tube 64 is arranged at an upper position at the back (arrow Y2 direction side) of the first receiving section 621 of the reagent container holder 62, and is configured to be moved in the vertical direction (Z direction) by an aspirating tube lifting mechanism 65 that holds the aspirating tube 64. As a result, the aspirating tube 64 is configured to enter the first storage section 210 (310) through the entrance section 212 (312) of the reagent container 200 (300) inserted at the back side of the reagent container holder 62, and to aspirate the reagent inside the reagent container 200 (300). In addition, the aspirating tube 64 is formed so that the tip end can penetrate (puncture) the seal member 213 (313) for sealing the opening 212a (312a) (see Figures 17 and 18) formed in the entrance section 212 (312) of the reagent container 200 (300). As described with reference to Figures 2, 5 and 6, the suction tube 64 constitutes one end of the liquid delivery tube 431, and the upper end of the suction tube 64 is connected to a flow path (not shown in Figures 19 to 21) leading to the liquid delivery section 430 and the chamber 420.

[0094] As shown in FIG. 19 and FIG. 20, the aspirating tube lifting mechanism 65 is configured to hold the aspirating tube 26 and the cover 63. The aspirating tube lifting mechanism 65 is engaged with grooves 611 and 612 provided in the chassis 61 so as to be movable in the vertical direction (Z direction). Thus, the aspirating tube lifting mechanism 65 is configured to integrally move the aspirating tube 26 in the vertical direction (Z direction) in conjunction with the opening and closing (up and down movement) of the cover 63. As shown in FIG. 19, when the cover 63 is arranged at the raised position P2, the aspirating tube 26 is arranged at the raised position P3 above the reagent container holding part 62 (outside the reagent container 200 (300) and the first receiving part 621). As shown in FIG. 21, when the cover 63 is arranged at the lowered position Q2, the aspirating tube 26 is arranged at the lowered position Q3 close to the inner bottom part immediately below the entrance part 212 (312) of the reagent container 200 (300).

[0095] The tip of the aspirating tube 26 inserted into the reagent container becomes the first end of the liquid delivery tube. The first end of the aspirating tube 26 is fixed at the above-mentioned lowered position Q3 while the reagent container is placed in the device. In other words, the first end of the aspirating tube 26 is fixed at the lowered position Q3 while measurements of multiple samples are performed using the reagent in the reagent container.

[0096] As shown in Figures 22 and 25, a large reagent container 200 (capacity approximately 100 mL) and a small reagent container 300 (capacity approximately 20 mL) are configured to be used according to the type of reagent to be contained therein. Each of the reagent containers 200 and 300 integrally includes a first storage section 210 (310) having an entrance section 212 (312) at the top into which the aspirating tube 26 can enter, and a second storage section 220 (320) continuous with the first storage section 210 (310). As shown in Figures 17 and 18, the first storage section 210 (310) is configured to be placed inside the first receiving section 621 with the reagent container 200 (300) set in the reagent container holder 62. As shown in Figures 17 and 18, the second accommodating section 220 (320) is configured to be disposed outside the first receiving section 621 with the reagent container 200 (300) set in the reagent-container holding section 62. As shown in Figures 23 and 26, a first reagent accommodating space 211 (311) is provided inside each first accommodating section 210 (310), and a second reagent accommodating space 221 (321) continuous with the first reagent accommodating space 211 (311) is provided inside each second accommodating section 220 (320).

[0097] The first storage section 210 (310) is a portion having a length L11, and the shape thereof is approximately common to the reagent containers 200 and 300. Since the shapes of the first storage sections 210 and 310 of the reagent containers 200 and 300 are approximately common, they can be set in the reagent container holding sections 62 (first receiving sections 621) of the holder sections 60a to 60e, which have the same shape.

[0098] Specifically, as shown in Figs. 17 and 18, each of the first storage sections 210 (310) has a constant width W21 that is slightly smaller than the width W11 of the first receiving section 621. Also, each of the first storage sections 210 (310) is provided with an entrance section 212 (312) at the end on the front side (the direction in which the first storage section 210 (310) is inserted into the reagent container holding section 62, the direction of the arrow Y2 in Figs. 17 and 18). Therefore, the reagent container 200 (300) is configured to be inserted from the entrance section 212 (312) side of the first storage section 210 (310) toward the back side of the reagent container holding section 62. This entrance section 212 (312) is provided so as to protrude upward from the outer upper surface 200b (300b) as shown in Figs. 23 and 26. As shown in Figures 22 and 25, the protruding entry portion 212 (312) is formed with an opening 212a (312a) that communicates with the inside of the first storage portion 210 (310). A seal member 213 (313) made of aluminum foil or the like is provided in the entry portion 212 (312) so as to close the opening 212a (312a), and is configured to seal the reagent container 200 (300). The outer diameter of the entry portion 212 (312) is equal to the width W21 of the first storage portion 210 (310), and the entry portion 212 (312) is formed so as to protrude continuously (flush) from the front surface of the first storage portion 210 (310).

[0099] 23 and 26, the reagent container 200 (300) is configured such that the inner bottom surface 200a (300a) is non-parallel to the outer top surface 200b (300b), and the distance between the inner bottom surface 200a (300a) and the outer top surface 200b (300b) increases toward the entrance portion 212 (312). In the second embodiment, the inner bottom surface 200a (300a) is configured to be an inclined surface inclined at an angle θ1 (approximately 10 degrees) with respect to the outer top surface 200b (300b). The reagent container 200 (300) has a bottom 200c (300c) that is approximately parallel to the outer upper surface 200b (300b) at the bottom surface directly below the entrance 212 (312), and the inclined surface 200d (300d) starts from the end of the bottom 200c (300c). As a result, when the reagent container 200 (300) is set at the set position Q1 shown in Fig. 20, the entrance 212 (312) is located at the uppermost position, and the bottom 200c (300c) directly below the entrance 212 (312) is located at the lowermost position.

[0100] Further, the outer upper surface 200b (300b) of the reagent container 200 (300) is provided with a protruding portion 230 (330) that protrudes upward (perpendicular to the outer upper surface 200b (300b)). The protruding portion 230 (330) has a plate-like shape with a length L1 extending in the longitudinal direction of the reagent container 200 (300), and is formed so as to have a protruding amount (protruding height) that is approximately equal to that of the entry portion 212 (312). The protruding portion 230 (330) is provided at a position near the end of the rear side (the side in the direction of the arrow Y1 in Figures 17 and 18) of each second storage portion 220 (320), and functions as a handle portion so that the user can easily set or remove the reagent container 200 (300).

[0101] On the other hand, the shape of the second accommodating section 220 (320) differs between the reagent container 200 and the reagent container 300. As shown in Fig. 23, the second accommodating section 220 of the large reagent container 200 integrally includes a first section 222 that is continuous with the first accommodating section 210 and whose width increases with increasing distance from the first accommodating section 210, and a second section 223 that has a constant width W22 that is larger than the width W21. Therefore, as shown in Fig. 23, in the reagent container 200, the second reagent accommodating space 221 inside the second accommodating section 220 is a reagent accommodating space that is provided continuously over both the first section 222 and the second section 223.

[0102] 23, the first section 222 is continuous with the second section 223 so as to expand the width of the first storage section 210 at an angle θ2 (approximately 60 degrees), and connects the first storage section 210 and the second section 223. In addition, the second section 223 has a width W22 larger than the width W21, so that the capacity of the second reagent storage space 221 can be secured as approximately 100 mL.

[0103] As shown in Figures 16 and 17, the intermediate receiving portion 622 of the reagent container holding portion 62 and the second receiving portion 623 which is continuous with the intermediate receiving portion 622 and has a width W12, respectively have shapes which correspond to the first portion 222 and the second portion 223 which has a width W22.

[0104] 26, the second storage section 320 of the small reagent container 300 has a constant width W21 that is the same as the width W21 of the first storage section 310. That is, in the small reagent container 300, the first storage section 310 and the second storage section 320 are formed to extend continuously in a straight line. The length L22 of the second storage section 320 is smaller than the length L21 of the second storage section 220 of the large reagent container 200. The small reagent container 300 is configured to have a total reagent capacity of about 20 mL by including the second storage section 320 having a width W21 and length L22 smaller than those of the second storage section 220 of the large reagent container 200.

[0105] As described above, the length L11 of the first storage section 310 is common to the reagent containers 200 and 300 because it is set in the first receiving section 621. Therefore, as shown in FIG. 18, for a small reagent container 300 in which the first storage section 310 and the second storage section 320 are continuous with the same width W21, the area stored in the first receiving section 621 is the first storage section 310, and the area disposed outside the first receiving section 621 is the second storage section 320.

[0106] Furthermore, unlike the large reagent container 200, the small reagent container 300 has a recess 340 on the inclined surface 300d of the outer bottom surface, which extends linearly along the longitudinal direction of the reagent container 300. When the inclined surface 300d is placed on a horizontal surface, this recess 340 allows the outer periphery of the recess 340 to be a contact point with the horizontal surface, so that even a reagent container 300 having a small width W21 can be stably stood upright.

[0107] As shown in Figs. 22 and 25, a label 250 (350) is attached to each reagent container 200 (300) on which the name of the reagent contained therein, the lot number of the reagent, the expiration date, an identification barcode, and the like are printed. The label 250 (350) is attached to the rear surface and at least one of the lateral sides of each reagent container 200 (300). A color indicating the type of reagent contained therein is applied to a part (a part corresponding to the rear surface of each reagent container 200 (300)) or the whole of the label 250 (350), so that the type of reagent can be identified by the color displayed on the label 250 (350). It is possible to check whether the reagent container 200 (300) is set in the correct holder section 60a to 60e by checking whether the colors of the label 250 (350) and the label 632 (see Fig. 15) attached to the cover 63 of the reagent container holder 60 match.

[0108] The reagent in the reagent container 200 (300) can be stored at the temperature of the environment where the analysis system 4002 is installed. The environmental temperature where the analysis system 4002 is installed is, for example, a temperature in the range of 20°C to 35°C. For example, at the environmental temperature, the reagent in the reagent container 200 (300) can be stored in the device in a state where performance can be guaranteed for several months (for example, 60 days, 90 days, or 120 days) after the reagent container 200 is installed in the device and the seal member 213 of the entrance part 212 is opened. The reagent contained in the reagent container 200 (300) contains, for example, multiple fluorescent dyes each corresponding to multiple wavelengths of light irradiated from the light source of the FCM detection unit 460. The compounds themselves that constitute the dye of these fluorescent dyes stain the cytoplasm, nucleic acid, and DNA of cells. As a method for staining cells in blood, for example, immunostaining using an antibody reagent containing a labeled antibody that specifically binds to a cell surface antigen (e.g., CD4 or CD25) is known, but such an antibody reagent needs to be cooled and stored in a refrigerator after being used for a certain period of time (e.g., 8 hours). In other words, the user needs to remove the antibody reagent from the device at regular intervals and return it to the refrigerator, and then set the antibody reagent in the device again when using it. In this respect, the reagent of this embodiment does not need to be refrigerated because the compound (fluorescent dye) that constitutes the dye itself stains cells and does not contain antibodies, and can be stored at the temperature of the device's operating environment. Therefore, in this embodiment, once the reagent container is placed in the device, it is not necessary to replace it until the reagent is used up, so long as the performance is guaranteed, and the frequency of reagent replacement is low, reducing the user's operating costs.

[0109] Third embodiment In this embodiment, the reagent container 200 is configured to suppress deterioration of the quality of a reagent containing multiple fluorescent dyes after installation in the measurement unit 400, and to enable continued use for a longer period of time while remaining attached to the measurement unit 400. One of the main means for suppressing deterioration is to prevent contact between the reagent and air. In order to prevent contact with air, the opening of the reagent container 200 is sealed when a liquid supply tube is inserted into the reagent container 200. The reagent container 200 includes a flexible bag-shaped reagent storage section so that the reagent can be supplied by the liquid supply tube even in a state in which contact between the air and the reagent is prevented by sealing the opening of the reagent container 200.

[0110] The storage period of the reagent is, for example, 75 days to one year after the reagent container 200 is installed in the measurement unit 400. The storage period is the length of time during which the measurement accuracy using the reagent by the measurement unit 400 can be guaranteed. In this embodiment, for example, the measurement accuracy is maintained for a period of 75 days to one year with the inside of the reagent storage section 10 sealed. Since the deterioration of the reagent can be suppressed by sealing the reagent storage section 10, the quality of the reagent can be stably maintained for a longer period. As a result, the frequency of replacement of the reagent (reagent container 200) containing multiple fluorescent dyes can be reduced, and the burden on the user regarding the replacement work can be reduced. The multiple fluorescent dyes may be contained in different reagent containers 200, or may be contained in one reagent container 200.

[0111] A reagent container 200 according to another embodiment will be described with reference to Figs. 28 and 29. The reagent container 200 is installed in the measurement unit 400 as shown in Fig. 28(A). As shown in Fig. 28(B), an aspirating tube 252 is inserted into the reagent container 200 from above in conjunction with a predetermined operation on the measurement unit 400. The aspirating tube 252 constitutes the first end of the above-mentioned liquid delivery tube. As shown in Fig. 28(B), the reagent container 200 contains the reagent 12 that is repeatedly aspirated in a state in which the aspirating tube 252 provided in the measurement unit 400 is inserted. The reagent container 200 includes a reagent storage section 10 and a frame 20. Hereinafter, the up-down direction is referred to as the Z direction. As shown in Fig. 29, the longitudinal direction of the reagent container 200 in the horizontal direction is referred to as the first direction A, and the lateral direction of the reagent container 200 is referred to as the second direction B.

[0112] The reagent containing section 10 contains a reagent 12 (see FIG. 29). As shown in FIG. 29, the reagent containing section 10 is a bag-shaped liquid containing member. The capacity of the reagent containing section 10 is, for example, 200 mL or more and 500 mL or less, or 20 mL or more and 100 mL or less.

[0113] The reagent containing section 10 is formed into a bag shape using a film-like material. The reagent containing section 10 is flexible and deformable. As shown in Fig. 30, the reagent containing section 10 is formed into a hollow bag shape by overlapping a plurality of sheet-like members and bonding the outer peripheries of the overlapped film materials. The bag shape may also be formed by bonding the inner surfaces of the outer peripheries of a single folded film material.

[0114] The reagent storage section 10 is made of a laminated film material 11 having gas barrier properties and light-shielding properties. Gas barrier properties are properties that make it difficult for gas to pass through. In this specification, gas barrier properties refer to the difficulty of air, particularly oxygen, passing through. Light-shielding properties are properties that make it difficult for light to pass through. This makes it possible to prevent the reagent 12 contained in the reagent container 200 from being deteriorated by external air and the reagent 12 contained in the reagent container 200 from being deteriorated by external light such as sunlight. As a result, deterioration of the reagent can be effectively prevented over a long period of time.

[0115] The laminated film material 11 may include at least one base layer and at least one gas barrier layer. The laminated film material 11 may further include a protective layer that protects the outer surface of the gas barrier layer. The laminated film material 11 may have a light-shielding layer made of a light-shielding material. When a material having both gas barrier and light-shielding properties is used for the gas barrier layer, the gas barrier layer and the light-shielding layer may be the same layer. The number of layers of the laminated film material is 2 or more, but may be 3 to 9 or 10 or more, and is not particularly limited.

[0116] The laminated film material 11 used in the reagent containing section 10 may have the following structure, for example: (outside of bag) / nylon (15 μm) / aluminum foil (9 μm) / polyethylene (9 μm) / (inside of bag). In this structure, the nylon functions as a protective layer, the aluminum foil functions as a gas barrier layer and a light-shielding layer, and the polyethylene is a base material layer in the reagent containing section 10. The polyethylene base material layers in the reagent containing section 10 are joined together by thermal welding. The laminated film material 11 with this structure is a metal foil laminate film having a structure in which a gas barrier layer made of metal foil is laminated on a resin base material layer.

[0117] For example, a resin-based multi-layer barrier film, a coating-based film, a vapor deposition film, an organic-inorganic composite film, etc. can be used for the laminated structure film material 11. The resin-based multi-layer barrier film is a film having a structure in which gas barrier layers of resin materials are laminated. Examples of resin materials with excellent gas barrier properties include PVDC (polyvinylidene chloride), PVA (polyvinyl alcohol), and EVOH (ethylene-vinyl alcohol copolymer). The coating-based film is a film having a structure in which a gas barrier material is coated (formed) on a base layer. Examples of gas barrier materials to be formed include PVDC, PVA, and EVOH. The vapor deposition film is a film having a structure in which a gas barrier material is vapor-deposited on a base layer. Examples of vapor-deposited gas barrier materials include metals such as aluminum, or inorganic oxides such as alumina and silica. Examples of organic-inorganic composite films include laminated films having a structure in which a gas barrier layer of an organic material (resin material) and a gas barrier layer of an inorganic material are separately laminated, and films having a gas barrier layer in which an inorganic material is dispersed in an organic binder.

[0118] The frame 20 includes an opening 21a attached to the reagent containing section 10. The frame 20 is configured so that the inside of the reagent containing section 10 is sealed when the aspirating tube 252 is inserted from above in conjunction with a predetermined operation on the measurement unit 400.

[0119] 28, the reagent container 200 is installed and held in a predetermined installation position in the container holding section 251 of the measurement unit 400 by the user. With the reagent container 200 installed at the set position Ps of the container holding section 251, the aspirating tube 252 arranged above the reagent container 200 is inserted from above into the opening 21a. The set position Ps is a position where the opening 21a is located directly below the aspirating tube 252. The frame 20 is configured such that with the aspirating tube 252 inserted, the opening 21a is closed, thereby sealing the inside of the reagent storage section 10.

[0120] This eliminates the need for the user to insert the aspirating tube 252 into the opening 21a of the reagent container 200 before installing the reagent container 200, making it easy to insert the aspirating tube 252 into the reagent container 200. Furthermore, since the inside of the reagent containing section 10 is sealed when the aspirating tube 252 is inserted, the sealing can prevent the reagent in the reagent containing section 10 from coming into contact with the outside air. This can prevent deterioration of the reagent after installation in the measurement unit 400. Furthermore, by, for example, grasping the frame 20 or installing it in the measurement unit 400, the installation work of the reagent container 200 to the measurement unit 400 can be easily performed even if the reagent containing section 10 has a soft bag-like shape.

[0121] In the case of rigid, non-deformable reagent containers, when reagent is aspirated in a sealed state, the internal pressure of the reagent container decreases as the reagent is reduced, and this becomes equilibrium with the suction pressure, making it impossible to aspirate the reagent. Therefore, the inside of the container must be left open to the atmosphere, and the reagent in the reagent container may deteriorate due to contact with air.

[0122] In contrast, in this embodiment, since the reagent storage unit 10 is provided in a deformable bag shape, the reagent storage unit 10 itself shrinks and deforms, thereby preventing a drop in the internal pressure, and as a result, the reagent 12 can be aspirated even in a sealed state. Therefore, the reagent container 100 is configured to seal the inside of the reagent storage unit 10 by suppressing the inflow of air into the opening 21a. By suppressing the inflow of air into the opening 21a, deterioration of the reagent 12 inside the reagent storage unit 10 due to contact with air can be suppressed. Since the deterioration of the reagent can be suppressed, the reagent in the reagent container 200 can be used for a longer period of time while the reagent container 200 is installed in the measurement unit 400. Therefore, by increasing the capacity of the reagent storage unit 10 and enabling a larger number of times of aspirating the reagent from one reagent container 200, the frequency of replacement of the reagent container 200 accompanying the operation of the measurement unit 400 can be reduced. For the user, this has the advantage of reducing the number of times the reagent container 200 needs to be replaced in a certain period of time.

[0123] Furthermore, in a configuration in which the aspirating tube 252 is inserted into the opening 21a by the operation of the measuring unit 400 rather than manually inserting the aspirating tube 252 by the user, positioning the opening 21a and preventing the opening 21a from shifting in position are important for reliably sealing the reagent containing unit 10 without damaging the reagent containing unit 10 with the aspirating tube 252. The reagent container 200 of this embodiment has a frame structure for reliably positioning the opening 21a and preventing the opening 21a from shifting in position when installed in the measuring unit 400.

[0124] As shown in FIG. 29, the frame 20 includes an opening 21 having an opening 21a, and a movement restricting portion 23 configured to abut against a part of the measurement unit 400 to restrict the movement of the opening 21a.

[0125] Movement of opening 21a can be suppressed by movement restricting portion 23 coming into contact with a part of measurement unit 400. This can suppress misalignment between aspirating tube 252 and opening 21a when aspirating tube 252 is inserted. In addition, since misalignment between aspirating tube 252 and opening 21a can be suppressed when aspirating tube 252 is inserted into reagent containing portion 10 through opening 21a, the inside of reagent containing portion 10 can be effectively maintained in a sealed state.

[0126] The opening 21 is a cylindrical portion in which an opening 21a is formed. The lower end of the opening 21a is connected to the inside of the reagent containing unit 10, and the upper end of the opening 21a is connected to the outside of the reagent containing unit 10. The opening 21 is provided in the upper part of the reagent containing unit 10.

[0127] Opening 21 has a sealing surface 21b that comes into contact with a sealing body 252a (see FIG. 28) provided on aspirating tube 252 to seal opening 21a. This makes it possible to easily and effectively seal reagent containing section 10 by the contact between sealing body 252a and sealing surface 21b.

[0128] The sealing surface 21b may be any one of the upper end surface of the opening 21, the inner peripheral surface of the opening 21, or the outer peripheral surface of the opening 21. In the example of Fig. 29, the sealing surface 21b is the annular upper end surface of the opening 21 surrounding the opening 21a (i.e., the edge of the opening 21a), and comes into vertical contact with a sealing body 252a (see Fig. 28(B)) provided on the suction tube 252.

[0129] For example, a removable cap can be attached to the opening 21. The reagent container 200 can be provided to a user with the opening 21a sealed with a cap, and can be installed in the measurement unit 400 after the cap is removed.

[0130] Also, for example, a puncturable sealing film can be welded to cover the opening 21a on the upper surface of the opening 21. The reagent container 200 is provided to a user with the opening 21a sealed by the sealing film, and after being placed in the measurement unit 400, the sealing film can be punctured by the suction tube 252 to open the reagent container.

[0131] 30, the opening 21 is formed in a mounting member 22 attached to the reagent containing section 10. The opening 21 is integrally formed with the mounting member 22 so as to protrude upward from the upper surface of the mounting member 22.

[0132] The outer periphery of the mounting member 22 is welded to the inner periphery of the reagent containing section 10. The mounting member 22 is provided at the upper end of the reagent containing section 10 so as to be sandwiched between the outer peripheries of the two laminated film materials 11 that constitute the reagent containing section 10, and is welded to the inner surfaces of each of the two laminated film materials 11. The space between the outer periphery of the mounting member 22 and the reagent containing section 10 is sealed by welding. This makes it possible to easily provide the mounting member 22, which has the rigidity required for coupling with the movement restricting section 23, in the flexible, bag-shaped reagent containing section 10, and also makes it easy to seal the space between the mounting member 22 and the reagent containing section 10.

[0133] The mounting member 22 has a supported portion 22a arranged at a horizontal distance from the opening 21. The mounting member 22 has an elongated hexagonal shape, with the opening 21 arranged at one end in the longitudinal direction of the mounting member 22 and the supported portion 22a arranged at the other end. The supported portion 22a is a protrusion formed integrally with the mounting member 22 so as to protrude upward from the upper surface of the mounting member 22. When viewed from the A direction, the supported portion 22a has a T-shape, with the base portion being thinner than the upper portion.

[0134] Here, opening 21 is located near the end of reagent containing unit 10 in the horizontal direction. Even when reagent containing unit 10 is in a bulging state due to containing reagent 12 (see FIG. 28), opening 21 of mounting member 22 is located relatively close to the inner surface of reagent containing unit 10. Therefore, if aspirating tube 252 is inserted at an angle to the central axis of opening 21, there is a possibility that the tip of aspirating tube 252 will come into contact with the inner surface of reagent containing unit 10.

[0135] Therefore, the opening 21 has an aspirating tube guide 21c that protrudes downward from the lower end of the opening 21a toward the inside of the reagent containing unit 10. The aspirating tube guide 21c is formed integrally with the mounting member 22, and has a shape in which the lower end of the cylindrical opening 21 is partially extended downward. The aspirating tube guide 21c is formed in a wall shape so as to separate the aspirating tube 252 inserted into the opening 21a from the inner surface of the reagent containing unit 10. This makes it possible to guide the tip of the aspirating tube 252 so as to prevent contact between the tip of the aspirating tube 252 and the inner surface of the reagent containing unit 10, even if the aspirating tube 252 is inserted into the inside of the reagent containing unit 10 at an angle with respect to the central axis of the opening 21a.

[0136] 29, the movement restricting part 23 has a function of positioning the opening 21a relative to the suction tube 252 of the measurement unit 400. In addition, the movement restricting part 23 has a function of preventing the opening 21a from shifting in position.

[0137] Movement restricting portion 23 is fixed to opening 21 so as to protrude horizontally from opening 21. This makes it possible to fix the relative positions of movement restricting portion 23 and opening 21a. Furthermore, the portion of movement restricting portion 23 protruding horizontally from opening 21 can easily realize a configuration in which movement is restricted by contacting a part of measurement unit 400, and the contact area required to prevent positional deviation of opening 21a can be easily obtained.

[0138] As shown in FIG. 30, the movement restriction part 23 has a generally flat plate shape. The movement restriction part 23 has a shape extending along a first direction A in a horizontal plane. The movement restriction part 23 has a rectangular shape including a pair of long sides extending in the first direction A and a pair of short sides extending in a second direction B. As described later, the reagent container 200 is installed in the container holding part 251 by inserting the reagent container 200 into the container holding part 251 of the measurement unit 400 along the first direction A, starting from the one end 23a side of the movement restriction part 23 where the opening 21 is arranged, as the leading end. The movement restriction part 23 is fixed to the opening 21 between the one end 23a and the center part of the movement restriction part 23 in the first direction A. That is, the opening 21 is arranged at a position closer to the one end 23a side of the movement restriction part 23 between the one end 23a and the other end 23b. As a result, by gripping the other end 23b side of the movement restricting part 23 and moving the reagent container 200 in the first direction A, the opening 21a formed in the opening 21 on the one end 23a side can be positioned directly below the aspirating tube 252. In this case, since the opening 21 is separated from the other end 23b of the movement restricting part 23, it is possible to prevent the user's fingers from coming into contact with the aspirating tube 252 when attaching the reagent container 200.

[0139] The movement restricting portion 23 is configured to restrict the horizontal movement of the opening 21a by abutting against a part of the measurement unit 400 in the horizontal direction. This allows the movement restricting portion 23 to suppress deviation of the horizontal positional relationship between the opening 21a and the suction tube 252. The movement restricting portion 23 includes a horizontal abutment surface 24 consisting of any one of the inner surface of a hole formed in the movement restricting portion 23, the inner surface of a notch formed in the movement restricting portion 23, and a side surface of the movement restricting portion 23. This allows the horizontal abutment surface 24 for restricting the horizontal movement of the opening 21a to be provided in the movement restricting portion 23 with a simple configuration.

[0140] The horizontal contact surface 24 includes a first surface 24a consisting of a side surface (i.e., a tip surface) formed at one end 23a of the movement restricting part 23 in the first direction A. As a result, when attaching the reagent container 200 to the measurement unit 400, both the alignment of the opening 21a in the first direction A and the restriction of movement can be easily achieved by simply moving the reagent container 200 in the first direction A until the one end 23a of the movement restricting part 23 abuts against the inner surface of the container holding part 251.

[0141] The horizontal contact surface 24 includes second surfaces 24b, 24c consisting of side surfaces extending along the first direction A of the movement restricting portion 23. The second surfaces 24b, 24c are a pair of side surfaces that form the longer sides of the movement restricting portion 23 in a plan view, and are provided on one side and the other side of the movement restricting portion 23 in the second direction B. This makes it possible to easily achieve both the alignment of the opening 21a in the second direction B and the restriction of movement.

[0142] As shown in FIG. 31, the movement restricting part 23 has a notch 23c formed on the side surface. The horizontal abutment surface 24 includes a third surface 24d formed of the inner surface of the notch 23c. The third surface 24d abuts against a part of the measurement unit 400 entering the notch 23c. This allows the movement restricting part 23 and the measurement unit 400 to engage with each other by the abutment of the third surface 24d in the notch 23c and a part of the measurement unit 400. This engagement can effectively prevent the position of the opening 21a from shifting. In addition, when the reagent container 200 is attached to the measurement unit 400, the movement restricting part 23 engages with a part of the measurement unit 400 and does not move, so that the user can feel that the reagent container 200 is installed in the correct position without visually confirming it.

[0143] The movement restricting portion 23 has a hole 23d formed on the upper surface of the movement restricting portion 23. The horizontal abutment surface 24 includes a fourth surface 24e formed of the inner surface of the hole 23d. The fourth surface 24e abuts against a part of the measurement unit 400 (for example, an entry member 281 described later, see FIG. 35) that enters the inside of the hole 23d. This allows the movement restricting portion 23 and the measurement unit 400 to engage with each other by a part of the measurement unit 400 entering the hole 23d formed in the movement restricting portion 23. The engagement can effectively prevent the position of the opening 21a from shifting. In FIG. 31, two holes 23d are provided. The two holes 23d are aligned along the second direction B. There may be only one hole 23d.

[0144] Movement restricting part 23 has guide part 23e that guides the movement of opening 21a when it is positioned below aspirating tube 252 of measuring unit 400. Guide part 23e extends along first direction A in a horizontal plane. Guide part 23e slides while in contact with a part of measuring unit 400, thereby aligning the movement direction of movement restricting part 23 with first direction A. This makes it possible to easily and accurately position opening 21a at an appropriate position where aspirating tube 252 can be inserted when reagent container 200 is placed in container holding part 251 of measuring unit 400.

[0145] Movement restricting portion 23 is configured to restrict vertical movement of opening 21a by abutting in the vertical direction (Z direction) against a part of measurement unit 400. This makes it possible to restrict vertical movement of opening 21a even in the case of reagent container 200 including a bag-shaped reagent containing portion 10 that is easily deformed when aspirating tube 252 is inserted into reagent containing portion 10 and when aspirating tube 252 is pulled out from reagent containing portion 10.

[0146] 31, the movement restricting part 23 has a gripping part 25 for gripping the movement restricting part 23 at the other end 23b of the movement restricting part 23 in the first direction A. This makes it easy to grip the other end 23b side of the movement restricting part 23. In addition, since the bag-shaped reagent storage part 10 is easily deformed and difficult to grip, providing the gripping part 25 on the movement restricting part 23 makes it easier to carry the reagent container 200.

[0147] 30, the movement restricting part 23 includes a fixing part 26 fixed to the opening 21, and a support part 27 that supports an upper part of the reagent containing part 10 at a position away from the opening 21. As a result, when the movement restricting part 23 is gripped by hand during carrying of the reagent container 200, the reagent containing part 10 can be supported so as to be suspended from the movement restricting part 23 at multiple points of the fixing part 26 and the support part 27. Since the weight of the reagent containing part 10 acts in a distributed manner at multiple points of the movement restricting part 23, the reagent containing part 10 can be stably supported even if the capacity of the reagent containing part 10 is large. Furthermore, the user does not have to grip the bag-shaped part, which is easily deformed, and this makes it easier to handle the reagent container 200.

[0148] As shown in Fig. 29, the movement restricting section 23 includes an information recording medium 28 on which information about the reagent is recorded. The information recording medium 28 is a radio frequency identifier (RFID) tag. This allows the measurement unit 400 to read information about the reagent from the information recording medium 28 when the reagent container 200 is attached to the measurement unit 400. When the information recording medium 28 is provided in a bag-shaped reagent containing section 10 that is easily deformed, it may be difficult to attach the information recording medium 28 or the information recording medium 28 may be difficult to read. However, by providing the information recording medium 28 in the movement restricting section 23, the information recording medium 28 can be easily provided and can be read satisfactorily.

[0149] As shown in Fig. 32, the reagent aspirating section 250 includes a plurality of reagent container holders each capable of holding one reagent container 200 (or small reagent container 300). The reagent aspirating section 250 includes, for example, a total of five reagent container holders, but for convenience, only three reagent container holders 250a, 250b, and 250c are shown in Fig. 32. The multiple reagent container holders 250a to 250c are arranged side by side in the horizontal direction.

[0150] Each of the reagent container holders 250a to 250c includes a lower chassis 255a and an upper chassis 255b, a container holding section 251, an aspirating tube 252, an operation section 253, and a moving mechanism 254 (see FIG. 35). The structure of the reagent container holder 250a will be described below as a representative of the reagent container holders 250a to 250c. The container holding section 251 is provided on the lower chassis 255a. The container holding section 251 is located at the bottom of the reagent container holder 250a. The container holding section 251 is configured to hold a reagent container 200 including a bag-shaped reagent containing section 10 for containing a reagent and a frame 20 including an opening 21a.

[0151] The container holding unit 251 has a box-shaped or cylindrical storage unit 260 capable of storing the reagent container 200 therein and having an inlet formed in the horizontal direction. The storage unit 260 has an inlet that opens at the end in the Y1 direction and is provided so as to extend from the inlet in the Y2 direction.

[0152] As shown in Figure 33, the storage section 260 includes a first insertion section 261 into which the reagent storage section 10 of the reagent container 200 is inserted, and a second insertion section 262 positioned above the first insertion section 261 and into which the movement control section 23 of the reagent container 200 is inserted.

[0153] The first insertion section 261 is a space defined by a pair of side surface portions 261a facing both side surfaces of the reagent containing section 10 in the width direction (X direction) and a bottom surface portion 261b facing the lower surface of the reagent containing section 10 in the up-down direction (Z direction). An upper portion of the first insertion section 261 and the second insertion section 262 are connected by a connecting passage 263.

[0154] The connection passage 263 is a space for passing through the connecting portion (the portion where the mounting member 22 is disposed) of the reagent container 200 between the movement restricting portion 23 and the reagent containing portion 10 in the Y direction.

[0155] The second insertion section 262 is configured to extend from the entrance of the storage section 260 toward the set position Ps directly below the aspirating tube 252, and to guide the reagent container 200 to the set position Ps (see FIG. 34) by abutting against the movement restricting section 23. This allows the reagent container 200 to be guided from the entrance of the storage section 260 to the set position Ps using the movement restricting section 23. As a result, even if the reagent container 200 includes a reagent storage section 10 that is easily deformed, the reagent container 200 can be accurately guided to the set position Ps by a simple action of the user simply inserting the reagent container 200 from the entrance of the storage section 260. The second insertion section 262 has, for example, a convex cross-sectional shape. That is, the second insertion section 262 has a convex cross-sectional shape due to a wide lower portion into which the flat plate-shaped movement restricting section 23 is inserted and a narrow upper portion into which the opening 21 protruding above the movement restricting section 23 is inserted.

[0156] The reagent aspirating section 250 includes an abutting section 270. The abutting section 270 abuts against a movement restricting section 23 provided on the frame 20 of the reagent container 200 placed in the container holding section 251 to restrict movement of the opening 21a. This allows the abutting section 270 to abut against the movement restricting section 23, thereby preventing misalignment between the aspirating tube 252 and the opening 21a when the aspirating tube 252 is inserted. Furthermore, since misalignment between the aspirating tube 252 and the opening 21a can be prevented when the aspirating tube 252 is inserted into the reagent containing section 10 through the opening 21a, the inside of the reagent containing section 10 can be effectively maintained in a sealed state.

[0157] The abutment portion 270 abuts against the movement restricting portion 23 provided on the frame 20 of the reagent container 200 placed in the container holding portion 251 to restrict the movement of the opening 21a. The abutment portion 270 abuts against the movement restricting portion 23, thereby making it possible to suppress misalignment between the aspirating tube 252 and the opening 21a when the aspirating tube 252 is inserted. Furthermore, since it is possible to suppress misalignment between the aspirating tube 252 and the opening 21a when the aspirating tube 252 is inserted into the inside of the reagent containing portion 10 from the opening 21a, it is possible to effectively maintain a sealed state inside the reagent containing portion 10.

[0158] The contact portion 270 is configured to restrict movement of the opening 21a by coming into contact with the movement restricting portion 23 fixed to the opening 21a of the reagent container 200. This fixes the relative position between the movement restricting portion 23 and the opening 21a, so that by bringing the contact portion 270 into contact with the movement restricting portion 23, the positioning and movement restriction of the opening 21a can be performed reliably and accurately.

[0159] At least a portion of the contact portion 270 is provided in the storage portion 260. With this, by inserting the reagent container 200 into the storage portion 260 from the inlet, the contact portion 270 and the movement restricting portion 23 come into contact with each other, and the movement of the opening 21a inside the storage portion 260 can be restricted.

[0160] The abutment portion 270 is disposed in the second insertion portion 262. Thus, by providing the abutment portion 270 in the second insertion portion 262 corresponding to the movement restricting portion 23 of the reagent container 200, the abutment portion 270 and the movement restricting portion 23 can be abutted in a preset relative positional relationship even in a reagent container 200 including a reagent storage portion 10 that is easily deformed. As a result, the positioning of the opening 21a and the restriction of movement can be performed accurately.

[0161] The abutment portion 270 is configured to abut against any of a hole formed in the movement restricting portion 23, a notch formed in the movement restricting portion 23, and a side surface of the movement restricting portion 23. This makes it possible to abut the abutment portion 270 against the movement restricting portion 23 and effectively restrict the horizontal movement of the opening 21a with a simple configuration.

[0162] When suction tube 252 moves downward, entering member 281 also moves downward together with suction tube 252 and enters inside hole 23d of movement restricting portion 23. When entering member 281 enters hole 23d, movement of movement restricting portion 23 is restricted.

[0163] The abutment portion 270 is configured to restrict the vertical movement of the opening 21a by vertically abutting against the movement restricting portion 23. This makes it possible to restrict the vertical movement of the opening 21a even in the case of a reagent container 200 including a bag-shaped reagent containing portion 10 that is easily deformed, when the aspirating tube 252 is inserted into the reagent containing portion 10 and when the aspirating tube 252 is pulled out from the reagent containing portion 10.

[0164] 35, the aspirating tube 252 is disposed with its tip facing downward at a position above the innermost part (the end part in the Y2 direction) of the storage section 260 of the container holding section 251. The aspirating tube 252 is configured to be held by a moving mechanism 254 and moved in the vertical direction (Z direction) by the moving mechanism 254. As a result, the aspirating tube 252 is configured to enter the inside of the reagent storage section 10 (110) through the opening 21a of the reagent container 200 (200) inserted at the inner side of the container holding section 251 and to aspirate the reagent inside the reagent container 200 (200).

[0165] The aspirating tube 252 has a seal 252a that seals the opening 21a by coming into contact with the opening 21a when the aspirating tube 252 is inserted into the opening 21a. This allows the reagent containing portion 10 to be easily and effectively sealed by the seal 252a.

[0166] Sealing body 252a is provided so as to surround the outer periphery of aspirating tube 252, and is configured to elastically deform upon contact with the edge of opening 21a (i.e., sealing surface 21b of opening 21) to seal opening 21a. This allows reagent containing section 10 to be securely sealed by sealing body 252a by utilizing the downward movement of aspirating tube 252 when inserted into opening 21a.

[0167] The sealing body 252a is made of an elastically deformable rubber material. In addition, a biasing member 252b for biasing the sealing body 252a toward the opening 21a is provided between the sealing body 252a and the suction tube holder 254a that holds the suction tube 252. The biasing member 252b is made of a compression spring.

[0168] The moving mechanism 254 holds the aspirating tube 252 so as to be movable in the vertical direction (Z direction) between the raised position P1 and the lowered position P2. The moving mechanism 254 includes an aspirating tube holder 254a and a linear motion mechanism composed of a linear rail 254b and a fixed slider 254c. The aspirating tube holder 254a is attached with the aspirating tube 252, and the aspirating tube holder 254a is connected to the upper end of the linear rail 254b via a connecting part 254d extending in the Y direction. The linear rail 254b is disposed on the front surface (the side surface on the Y1 direction side) of the reagent container holder 250a and extends in the Z direction. The fixed slider 254c is fixed to the lower chassis 255a and holds the linear rail 254b so as to be movable along the Z direction. This allows the linear rail 254b to move in the Z direction relative to the fixed slider 254c. As the linear rail 254b moves in the Z direction, the suction tube 252 moves in the Z direction together with the linear rail 254b.

[0169] In this manner, the moving mechanism 254 is configured to support the aspirating tube 252 movably in the vertical direction, and to move the aspirating tube 252 in conjunction with a predetermined operation on the operation unit 253, thereby inserting the aspirating tube 252 into the reagent container 200 held in the container holding unit 251 and retracting the aspirating tube 252 out of the reagent container 200. This allows the aspirating tube 252 to insert into the reagent container 200 and retract out of the reagent container 200 simply by moving the aspirating tube 252 up and down. The smaller the degree of freedom in the movement direction of the aspirating tube 252, the more accurately and reproducibly the aspirating tube 252 can be operated, and therefore the operation of sealing the reagent containing unit 10 when the aspirating tube 252 is inserted can be performed more accurately by simple vertical movement.

[0170] 35, the movement mechanism 254 holds the operation unit 253 so that the aspirating tube 252 and the operation unit 253 move in conjunction with each other. The operation unit 253 is provided on the front side (the side in the Y1 direction) of the reagent container holder 250a, and the rear side (the Y2 direction side) of the operation unit 253 is attached to a linear rail 254b. This causes the linear rail 254b, the aspirating tube 252, and the operation unit 253 to move integrally in the Z direction.

[0171] The operation unit 253 is disposed on the front surface (the side in the direction of the arrow Y1) of each of the reagent container holders 250a to 250c (lower chassis 255a) as shown in Fig. 32. The operation unit 253 is configured as an openable and closable cover that covers the front surface of the container holding unit 251 of the reagent container holder 250a.

[0172] The operation unit 253 is configured so that the user can hold and move it with his / her hand. The operation unit 253 is formed with an operation grip portion 253a that protrudes forward (in the Y1 direction) from the front surface of the operation unit 253. The user can move the operation unit 253 in the Z direction by holding the operation grip portion 253a and moving it in the Z direction.

[0173] (Fourth embodiment) In the analysis systems 4000 and 4001 described in the first and second embodiments, the analysis unit (analysis section) 300X may input the waveform of the first signal and the waveform of the second signal corresponding to the fluorescence emitted from the cells flowing in the flow cell, and classify the cells using a trained AI algorithm for at least a part of the first to third analyses based on the waveform of the signal corresponding to the fluorescence emitted from the various cells to be classified. As described above, the first signal is a signal composed of a plurality of signals corresponding to the side scattered light and side fluorescent light from the first light source, and the second signal is a signal composed of a plurality of signals corresponding to the forward scattered light, side scattered light, and side fluorescent light from the second light source. In addition, the first analysis is an analysis based on the first signal, the second analysis is an analysis based on the second signal, and the third analysis is an analysis based on the first signal and the second signal.

[0174] In this embodiment, in an analysis system 4000 (see FIG. 7) including an FCM detection unit 460 capable of irradiating a measurement sample with light of multiple wavelengths as described above, data acquired by a measurement unit 400 is analyzed by an AI algorithm. Since light of multiple wavelengths is irradiated onto the measurement sample, the measurement unit 400 detects multiple types of optical signals corresponding to each wavelength. Data acquired by A / D converting each of the detected multiple types of optical signals is analyzed by an AI algorithm. Analysis by the AI ​​algorithm is performed by an analysis unit 300X (see FIG. 10). The AI ​​algorithm is stored, for example, in a memory unit 3004 of the analysis unit 300X. A processor 3001 of the analysis unit 302X executes the analysis based on the AI ​​algorithm.

[0175] As in the example shown in FIG. 36, data acquired by the measurement unit 402 (hereinafter, sometimes referred to as "waveform data") is input to the AI ​​algorithm 50 (or 60). The AI ​​algorithm processes the input data to classify the type of cell corresponding to the input data. When a measurement sample is irradiated with light of a single wavelength, only data corresponding to the light of a single wavelength is obtained for a certain cell in the measurement sample. When a measurement sample is irradiated with light of multiple wavelengths, multiple types of data corresponding to the lights of multiple wavelengths are obtained for a certain cell in the measurement sample. Therefore, when light of multiple wavelengths is irradiated to the measurement sample, the amount of data input to the AI ​​algorithm is greater than when light of a single wavelength is irradiated to the measurement sample. By increasing the amount of data input to the AI ​​algorithm, the accuracy of cell classification by the AI ​​algorithm is improved.

[0176] A method for generating training data 75 (see FIG. 40) and a method for analyzing waveform data will be described using the examples shown in FIG. 37 to FIG. 40. In the following, an example will be described in which an FCM detection unit 460 having the configuration in FIG. 7 is used.

[0177] <Waveform data> Fig. 37 is a schematic diagram for explaining waveform data used in this analysis method (fourth embodiment). As shown in Fig. 37, when a specimen containing cells (components) C is flowed through a flow cell FC and the cells C flowing through the flow cell FC are irradiated with light L1 of a first wavelength and light L2 of a second wavelength, forward scattered light (FSC) is generated forward with respect to the traveling direction of the light. First side scattered light (SSC-1) corresponding to the light of the first wavelength and first side fluorescence (SFL-1) excited by the light of the first wavelength are generated laterally with respect to the traveling direction of the light. In addition, second side scattered light (SSC-2) corresponding to the light of the second wavelength and second side fluorescence (SFL-2) excited by the light of the second wavelength are generated laterally with respect to the traveling direction of the light.

[0178] The forward scattered light (FSC) is received by the forward scattered light receiving element 4116, and a signal according to the amount of received light is output. The first side scattered light (SSC-1) is received by the side scattered light receiving element 4121a, and a signal according to the amount of received light is output. The first side fluorescent light (SFL-1) is received by the side fluorescent light receiving element 4122a, and a signal according to the amount of received light is output. The second side scattered light (SSC-2) is received by the side scattered light receiving element 4121b, and a signal according to the amount of received light is output. The second side fluorescent light (SFL-2) is received by the side fluorescent light receiving element 4122b, and a signal according to the amount of received light is output. As a result, an analog signal indicating the change in signal over time is output from each light receiving element. An analog signal corresponding to forward scattered light (FSC-1) is called the "forward scattered light signal," an analog signal corresponding to first side scattered light (SSC-1) is called the "first side scattered light signal," an analog signal corresponding to first side fluorescence (SFL-1) is called the "first fluorescence signal," an analog signal corresponding to second side scattered light (SSC-2) is called the "second side scattered light signal," and an analog signal corresponding to second side fluorescence (SFL-2) is called the "second fluorescence signal." One pulse of each analog signal corresponds to one component (e.g. one cell).

[0179] The analog signal is input to the A / D conversion unit 481a (see FIG. 37) via the analog processing unit 480 and converted into a digital signal. FIG. 38 is a diagram showing the conversion into a digital signal by the A / D conversion unit. Here, for the sake of simplicity, the diagram shows an analog signal being directly input to the A / D conversion unit 481a. As shown in FIG. 38, the A / D conversion unit 481a samples the forward scattered light signal, the first side scattered light signal, the first side fluorescent light signal, the second side scattered light signal, and the second side fluorescent light signal, starting from the point when the level of the forward scattered light signal reaches a level set as a predetermined threshold, among the analog signals input from each light receiving element. The A / D conversion unit 481a samples each analog signal at a predetermined sampling rate (for example, sampling of 1024 points at 10 nanosecond intervals, sampling of 128 points at 80 nanosecond intervals, or sampling of 64 points at 160 nanosecond intervals, etc.).

[0180] FIG. 39 is a diagram showing waveform data obtained by sampling. By sampling, matrix data (which may be called one-dimensional "array data") having values ​​digitally indicating analog signal levels at multiple time points as elements is obtained as waveform data corresponding to one component. In this way, the A / D conversion unit 481a generates a digital signal of forward scattered light, a digital signal of first side scattered light, a digital signal of first side fluorescent light, a digital signal of second side scattered light, and a digital signal of second side fluorescent light, which correspond to one component. The A / D conversion is repeated until the number of digitally converted cells reaches a predetermined number, or until a predetermined time has elapsed since the specimen began to flow through the flow cell 4113. As a result, as shown in FIG. 39, a digital signal consisting of waveform data of N components (cells C) contained in one specimen is obtained. A set of sampling data for each component (a set of 1024 digital values ​​every 10 nanoseconds from t=0 ns to t=10240 ns in the example of FIG. 39) may be called waveform data, and a set of waveform data obtained from one specimen may be called a digital signal. Waveform data is acquired by irradiating the flow cell 4113 with light of multiple wavelengths, and thus multiple waveform data corresponding to each wavelength are acquired. For example, in the example shown in Fig. 37, waveform data corresponding to first side scattered light and first side fluorescent light corresponding to light of a first wavelength (e.g., 405 nm), and waveform data corresponding to forward scattered light, second side scattered light, and second side fluorescent light corresponding to light of a second wavelength (e.g., 638 nm) are acquired. Side fluorescent light corresponding to light of the first wavelength is acquired based on a first fluorescent dye, and side fluorescent light corresponding to light of the second wavelength is acquired based on a second fluorescent dye.

[0181] An index for identifying each component may be assigned to each piece of waveform data generated by the A / D conversion unit 481a. For example, an integer from 1 to N is assigned to the index in the order of the generated waveform data, and the same index is assigned to the waveform data of the forward scattered light, the waveform data of the first / second side scattered light, and the waveform data of the first / second side fluorescent light obtained from the same component. Since each piece of waveform data corresponds to one component, the index corresponds to the measured component. By assigning the same index to the waveform data corresponding to the same component, the deep learning algorithm described later can analyze the waveform data of the forward scattered light, the waveform data of the first / second side scattered light, and the waveform data of the first / second fluorescent light corresponding to each component as one set, and classify the type of component.

[0182] <Generating training data> Fig. 40 is a schematic diagram showing an example of a method for generating training data used to train a deep learning algorithm for determining the type of component in a specimen. As shown in Fig. 40, the training data 75 is waveform data generated based on an analog signal 70a of forward scattered light (FSC), an analog signal 70b of first side scattered light (SSC-1), an analog signal 70c of first side fluorescent light (SFL-1), an analog signal 70d of second side scattered light (SSC-2), and an analog signal 70e of first side fluorescent light (SFL-2) obtained by measuring the specimen using a flow cytometer (measurement unit 400, see Fig. 7) for the components in the specimen. The method for acquiring the waveform data is as described above.

[0183] The training data 75 may be, for example, waveform data of a component determined to be highly likely to be a specific type as a result of measuring a sample with a flow cytometer and analyzing the components contained in the sample by computation. Hereinafter, an example will be described using an analysis system 4000 (see FIG. 4) as a blood cell counting device for analyzing blood samples. A blood sample is measured with a flow cytometer (measurement unit 400), and waveform data of forward scattered light, first / second side scattered light, and first / second side fluorescent light of each component contained in the sample are stored. Based on the first / second side scattered light intensity (pulse height of the side scattered light signal) and the first / second side fluorescent light intensity (pulse height of the side fluorescent light signal), the cells in the sample are classified into groups of neutrophils, lymphocytes, monocytes, eosinophils, basophils, immature granulocytes, and abnormal cells. The training data 75 is obtained by adding a label value corresponding to the classified cell type to the waveform data of the cell. For example, the mode, average or median of the side scattered light intensity and side fluorescent light intensity of cells contained in a population of neutrophils is determined, representative cells are identified based on these values, and the label value "1" corresponding to neutrophils is assigned to the waveform data of these cells, thereby obtaining training data 75. The method of generating training data is not limited to this, and training data may be obtained, for example, by collecting only specific cells using a cell sorter, measuring these cells using a flow cytometer, and assigning the label value of the cells to the obtained waveform data.

[0184] The analog signals 70a-70e respectively indicate the forward scattered light signal, the first / second side scattered light signal, and the first / second side fluorescent light signal when neutrophils are measured by the flow cytometer. When these analog signals are A / D converted as described above, waveform data 72a of the forward scattered light signal, waveform data 72b of the first side scattered light signal, waveform data 72c of the first side fluorescent light signal, waveform data 72d of the second side scattered light signal, and waveform data 72e of the second side fluorescent light signal are obtained. Adjacent cells within each of the waveform data 72a-72e store signal levels at intervals corresponding to the sampling rate, for example, at intervals of 10 nanoseconds. Each of the waveform data 72a to 72e is combined with a label value 77 indicating the type of cell from which the data is derived, and five waveform data corresponding to each cell, in other words, five signal intensities (signal intensity of forward scattered light, signal intensity of first / second side scattered light, and signal intensity of first / second side fluorescent light) are input to the deep learning algorithm (neural network 50) as training data 75 to form a set. In the example of FIG. 40, since the cell from which the training data 75 is derived is a neutrophil, a label value 77 of "1" indicating that the data is a neutrophil is assigned to each of the waveform data 72a to 72e, and the training data 75 is generated. An example of the label value 77 is shown in FIG. 41. Since the training data 75 is generated for each cell type, a label value 77 that differs depending on the cell type is assigned.

[0185] <Overview of deep learning> An overview of neural network training will be described with reference to FIG. 40 as an example. The neural network 50 is, for example, a convolutional neural network having a convolutional layer. The number of nodes in the input layer 50a of the neural network 50 corresponds to the number of elements of the array included in the waveform data of the input training data 75. The number of elements of the array is equal to the total number of elements of the waveform data 72a-72e of the forward scattered light, the first / second side scattered light, and the first / second side fluorescent light corresponding to one component. In the example of FIG. 40, each of the waveform data 72a-72e includes 1024 elements. Therefore, the number of nodes in the input layer 50a is 1024×5=5120. Each of the waveform data 72a-72e is input to the input layer 50a of the neural network 50. The label value 77 of each waveform data of the training data 75 is input to the output layer 50b of the neural network to train the neural network 50. The reference symbol 50c in FIG. 40 indicates an intermediate layer.

[0186] <How to analyze waveform data> FIG. 42 shows an example of a method for analyzing waveform data of components in a specimen. In the method for analyzing waveform data shown in FIG. 42, analysis data 85 consisting of waveform data obtained by the above-mentioned method is generated from an analog signal 80a of forward scattered light, an analog signal 80b of first side scattered light, an analog signal 80c of first side fluorescent light, an analog signal 80d of second side scattered light, and an analog signal 80e of second side fluorescent light acquired by a flow cytometer (measurement unit 400, see FIG. 7) from the component to be analyzed. Since the waveform data is acquired by irradiating light of multiple wavelengths onto the flow cell 4113 (see FIG. 37), multiple waveform data corresponding to each wavelength are acquired. For example, waveform data corresponding to the first side scattered light and the first side fluorescent light corresponding to the light of a first wavelength (e.g., 405 nm), and waveform data corresponding to the forward scattered light, the second side scattered light, and the second side fluorescent light corresponding to the light of a second wavelength (e.g., 638 nm) are acquired. Side fluorescence corresponding to the light of the first wavelength is obtained based on a first fluorescent dye, and side fluorescence corresponding to the light of the second wavelength is obtained based on a second fluorescent dye.

[0187] It is preferable that at least the acquisition conditions are the same for the analysis data 85 and the training data 75. The acquisition conditions include conditions for measuring components in a specimen by a flow cytometer, such as preparation conditions for the measurement sample, the flow rate when the measurement sample is flowed through a flow cell, the intensity of light irradiated onto the flow cell, the amplification factor (gain) of the light receiving element that receives scattered light and fluorescent light, etc. The acquisition conditions also include the sampling rate when analog signals are A / D converted.

[0188] When the components to be analyzed flow through the flow cell 4113, an analog signal 80a of forward scattered light, an analog signal 80b of first side scattered light, an analog signal 80c of first side fluorescence, an analog signal 80d of second side scattered light, and an analog signal 80e of second side fluorescence are obtained. When these analog signals 80a to 80e are A / D converted as described above, the time points at which the signal intensities were acquired are synchronized for each component, and waveform data 82a of the forward scattered light signal, waveform data 82b of the first side scattered light signal, waveform data 82c of the first side fluorescence signal, waveform data 82d of the second side scattered light signal, and waveform data 82e of the second side fluorescence signal are obtained. Each waveform data 82a to 82e is combined so that the data of the signal intensities of each component (signal intensity of forward scattered light, signal intensity of first / second side scattered light, and signal intensity of first / second side fluorescence) are combined into a set, and are input to the deep learning algorithm 51 as analysis data 85. In the example of FIG. 42, a data set made up of five pieces of waveform data 82a, 82b, 82c, 82d, and 82e is input.

[0189] When the analysis data 85 is input to the input layer 51a of the neural network 51 constituting the trained deep learning algorithm 51, the analysis result 83 is output from the output layer 51b as classification information on the type of the component corresponding to the analysis data 85. The reference symbol 51c in FIG. 42 indicates an intermediate layer. The classification information on the type of the component is, for example, the probability that each component belongs to each of a plurality of cell types. Furthermore, it may be determined that the component of the analysis target from which the analysis data 85 was obtained belongs to the classification with the highest value among these probabilities, and a label value 82 or the like that is an identifier representing the type may be included in the analysis result 83. The analysis result 83 may be data in which the label value is replaced with information indicating the type (for example, a character string) in addition to the label value itself. FIG. 42 shows an example in which the deep learning algorithm 51 outputs the label value "1" to which the component of the analysis target from which the analysis data 85 was obtained had the highest probability of belonging based on the analysis data 85, and further, character data of "neutrophil" corresponding to this label value is output as the analysis result 83. The label value may be output by the deep learning algorithm 51, or another computer program may output the most preferred label value based on the probability calculated by the deep learning algorithm 51.

[0190] Figure 43 shows an example of the configuration of another sample preparation unit 441 having a different configuration from the sample preparation unit 440 shown in Figure 12. In Figure 43, the same elements as those in Figure 12 are denoted by the same reference numerals.

[0191] The example of FIG. 43 shows that the configuration of the measurement channel of the sample preparation unit 440 can be changed by performing an analysis using a deep learning algorithm (AI algorithm). In the example of FIG. 43, a measurement channel (WNR) for counting white blood cells, nucleated red blood cells, and basophils is replaced with a measurement channel (WDF) for classifying white blood cells. That is, in the example of FIG. 43, the WNR channel is replaced with a WDF channel, so that the measurement unit 400 (see FIG. 7) is configured to include multiple WDF channels. In this example, the measurement items of the WNR channel, that is, the counting of white blood cells, the counting of nucleated red blood cells, and the counting of basophils, are obtained by analyzing the waveform data obtained by the measurement using the WDF channel with the AI ​​algorithm 51. For example, by having the AI ​​algorithm 51 learn so that white blood cells, nucleated red blood cells, and basophils can be classified from the waveform data obtained by the measurement using the WDF channel, the classification of white blood cells, nucleated red blood cells, and basophils becomes possible from the waveform data of the WDF channel. For example, by having the AI ​​algorithm 51 learn data corresponding to white blood cells, nucleated red blood cells, and basophils from waveform data measured in the WDF channel as training data, an AI algorithm 51 capable of classifying white blood cells, nucleated red blood cells, and basophils from waveform data measured in the WDF channel is generated. In the configuration example of Fig. 43, for example, measurements of different specimens are performed in parallel in multiple WDF channels. For example, sample preparation of different specimens is performed in parallel in each chamber of multiple WDF channels.

[0192] As shown in FIG. 43, the analysis of a specific measurement channel can be replaced by another measurement channel by replacing the analysis of the specific measurement channel with AI analysis of data from another measurement channel. This allows the number of measurement channels to be increased without increasing the total number of measurement channels provided in the analysis system 4002 (see FIG. 7). In the example of FIG. 43, by replacing the WNR channel with a WDF channel, it is possible to increase the number of WDF channels without increasing the total number of measurement channels provided in the analysis system 4002. By increasing the number of WDF channels, different samples can be measured in parallel in each of the multiple WDF channels. Parallel measurement improves the throughput of measurement by the WDF channels. The example of FIG. 43 has a remarkable effect of also improving the throughput of sample processing.

[0193] In the case of replacing a measurement channel with another measurement channel by utilizing an AI algorithm, it is required to analyze the measurement items of the measurement channel to be replaced (nucleated red blood cells and basophils in the WNR channel in the example of FIG. 12) based on data measured in the other measurement channel (WDF channel in the example of FIG. 12). By analyzing the data measured in the other measurement channel with an AI algorithm, it becomes possible to classify the measurement items of the measurement channel to be replaced. In this embodiment, multiple types of optical signals corresponding to multiple wavelengths of light are analyzed by the AI ​​algorithm. By increasing the amount of information using data corresponding to multiple wavelengths of light, the analysis accuracy by AI is improved. With the improvement in analysis accuracy, it becomes possible to classify the measurement items of the measurement channel to be replaced (nucleated red blood cells and basophils in the WNR channel in the example of FIG. 12) even if it is data of the other measurement channel (for example, the WDF channel).

[0194] In the analysis method (first example) in the example of FIG. 43, for example, by AI analysis of waveform data obtained by measurement using the WDF channel, in addition to classifying and counting NRBC (nucleated red blood cells) and BASO (basophils), classifying and counting white blood cells other than BASO (eosinophils, neutrophils, lymphocytes, monocytes) and counting are also performed. In this example, the AI ​​algorithm 51 is trained so that it can classify NRBC, BASO, and white blood cells other than BASO (eosinophils, neutrophils, lymphocytes, monocytes) using waveform data. By using such an AI algorithm 51, it is possible to replace the WNR channel with the WDF channel. These analyses are performed by the analysis unit 302X (see FIG. 7) as follows.

[0195] FIG. 44 is a flowchart showing an operation example (first operation example) of the present analysis method. As shown in FIG. 44, in the first operation example, the analysis unit 302X (see FIG. 7) acquires data corresponding to an optical signal detected in the WDF channel (step S0). Next, the analysis unit 302X analyzes this data with the AI ​​algorithm 51 (step S1). The analysis unit 302X analyzes the data obtained by measurement in the WDF channel with the AI ​​algorithm 51, thereby classifying basophils and nucleated red blood cells, which were analyzed in the WDF channel, in addition to classifying monocytes, lymphocytes, neutrophils, and eosinophils. Next, the analysis unit 302X provides an analysis result of the data in the WDF channel (step S2).

[0196] In another example (second operation example) of the analysis method shown in FIG. 45, for example, classification and counting of NRBC and classification and counting of BASO are performed by AI analysis of waveform data obtained by measurement using a WDF channel. Waveform data corresponding to cells not classified as NRBC or BASO by the AI ​​algorithm 51 is analyzed by a scattergram, and classification and counting of eosinophils, neutrophils, lymphocytes, and monocytes are performed. In this example, the AI ​​algorithm 51 is trained to classify, for example, from waveform data into NRBC, BASO, and others. Waveform data corresponding to cells classified as other than NRBC and BASO by the AI ​​algorithm 51 in this example is analyzed by a scattergram. For example, peak values ​​of waveform data corresponding to cells classified as other than NRBC and BASO are extracted, and cell types are classified based on a two-dimensional graph generated from peak values ​​corresponding to side fluorescent signals and peak values ​​corresponding to side scattered light. For example, based on a scattergram, cells are classified as eosinophils, neutrophils, lymphocytes, monocytes, and others. Cells classified as other than eosinophils, neutrophils, lymphocytes, and monocytes in the analysis based on the scattergram are classified as, for example, debris. These analyses are performed by the analysis unit 302X (see FIG. 7) as follows.

[0197] FIG. 45 is another flowchart showing an operation example (second operation example) of the present analysis method. As shown in FIG. 45, the analysis unit 302X acquires data corresponding to an optical signal detected in the WDF channel (step S10). Next, the analysis unit 302X analyzes the data with the AI ​​algorithm 51 (step S11). Next, the analysis unit 302X identifies data corresponding to cells other than NRBC and BASO (step S12). Next, the analysis unit 302X analyzes the identified data with a scattergram (step S13). Next, the analysis unit 302X provides an analysis result of the data of the WDF channel (step S14).

[0198] In yet another example (third example) of the analysis method shown in FIG. 46, for example, classification and counting of lymphocytes, classification and counting of monocytes, classification and counting of eosinophils, and classification and counting of neutrophils / basophils are performed by scattergram analysis of waveform data obtained by measurement using a WDF channel. In classification and counting of neutrophils / basophils, for example, cells classified as either neutrophils or basophils are counted. Waveform data corresponding to cells not classified as lymphocytes, monocytes, eosinophils, or neutrophils / basophils, and cells classified as either neutrophils or basophils, are analyzed by AI algorithm 51 (see FIG. 42). For example, the waveform data is classified by AI algorithm 51 into NRBC, basophils (BASO), and others. For example, the count of cells classified as BASO by the AI ​​algorithm 60 is subtracted from the count results of cells classified as either neutrophils or basophils in the scattergram analysis to calculate the count of neutrophils and the count of basophils, respectively. Cells that are not classified as either NRBC or BASO in the AI ​​analysis are classified as, for example, debris. These analyses are performed by the analysis unit 302X (see FIG. 7) as follows.

[0199] FIG. 46 is a flowchart showing an operation example (third operation example) of the present analysis method. As shown in FIG. 46, the analysis unit 302X acquires data corresponding to an optical signal detected in the WDF channel (step S20). Next, the analysis unit 302X analyzes the data in a scattergram (step S21). Next, the analysis unit 302X identifies data corresponding to (1) cells not classified as lymphocytes, monocytes, eosinophils, or neutrophils / basophils, and (2) cells classified as neutrophils / basophils (step S22). Next, the analysis unit 302X analyzes the identified data with the AI ​​algorithm 60 (step S23). Next, the analysis unit 302X provides an analysis result of the data of the WDF channel (step S24).

[0200] Fifth embodiment A fifth embodiment discloses a sample analysis method in a sample analysis system (including the analysis systems of the first to fourth embodiments) including a host processor and a parallel processing processor, comprising: acquiring data regarding each of the components in a sample based on control by the host processor, performing parallel processing on the data in the parallel processing processor, and generating information regarding the type of each of the components based on the results of the parallel processing.

[0201] According to this embodiment, even when analyzing huge amounts of data ranging from several hundred megabytes to several gigabytes per sample, the parallel processing processor provided separately from the host processor can execute processing related to the measured data in parallel. Therefore, even when processing huge amounts of data using a deep learning algorithm, data processing is completed within the sample analysis system. For example, there is no need to transmit data to an analysis server storing a deep learning algorithm via the Internet or an intranet. Therefore, according to this embodiment, there is no need to transmit large amounts of data from the sample analysis system to the analysis server and obtain the analysis results returned from the analysis server, and the processing capacity of the sample analysis system can be maintained while improving the classification accuracy of components in the sample.

[0202] FIG. 47 shows a configuration example of this embodiment. In FIG. 47, elements similar to those in FIG. 10 are given the same reference numerals. The analysis unit 303X in FIG. 47 includes a parallel processor capable of processing calculations by an AI algorithm instead of the master processor. By using a parallel processor suitable for processing matrix calculations executed by an AI algorithm (e.g., a deep learning algorithm), the TAT (Turn Around Time) required for AI analysis is improved. In the following description, descriptions of configurations and functions similar to those of the above-mentioned embodiment are omitted.

[0203] The analysis unit 302X shown in FIG. 47 may have a configuration in which the analysis section 301X is provided in the measurement unit 401 as shown in FIG.

[0204] The processor 3001 executes analysis processing of the waveform data by the deep learning algorithm 60 using the parallel processing processor 3002. That is, the processor 3001 is programmed to execute analysis processing of the waveform data according to the deep learning algorithm 60. Analysis software 3100 for analyzing data corresponding to components in a sample based on the deep learning algorithm 60 may be stored in the storage unit 3004. In this case, the processor 3001 executes analysis processing of data based on the deep learning algorithm 60 by executing the analysis software 3100 stored in the storage unit 3004. In this embodiment, for example, the AI ​​analysis is executed by the processor 3001 and the parallel processing processor 3002, and the computational analysis is executed by the processor 3001 without using the parallel processing processor 3002. The processor 3001 is, for example, a CPU (Central Processing Unit). The processor 3001 may be, for example, Intel's Core i9, Core i7, or Core i5, or AMD's Ryzen 9, Ryzen 7, Ryzen 5, or Ryzen 3.

[0205] The processor 3001 controls the parallel processing processor 3002. The parallel processing processor 3002 executes parallel processing related to, for example, matrix operations, according to the control by the processor 3001. In other words, the processor 3001 is a master processor of the parallel processing processor 3002, and the parallel processing processor 3002 is a slave processor of the processor 3001. The processor 3001 is also called a host processor or a main processor.

[0206] The parallel processing processor 3002 executes a plurality of arithmetic processes in parallel, which are at least a part of the process related to the analysis of the waveform data. The parallel processing processor 3002 is, for example, a graphics processing unit (GPU), a field programmable gate array (FPGA), or an application specific integrated circuit (ASIC). When the parallel processing processor 3002 is an FPGA, the parallel processing processor 3002 may be programmed in advance with arithmetic processes related to the trained deep learning algorithm 60, for example. When the parallel processing processor 3002 is an ASIC, the parallel processing processor 3002 may be pre-programmed with a circuit for executing arithmetic processes related to the trained deep learning algorithm 60, for example, or may be built-in with a programmable module in addition to such a built-in circuit. The parallel processing processor 3002 may be, for example, GeForce, Quadro, TITAN, Jetson, or the like manufactured by NVIDIA. For the Jetson series, for example, Jetson Nano, Jetson Tx2, Jetson Xavier, or Jetson AGX Xavier is used.

[0207] The processor 3001 executes, for example, calculation processing related to the control of the measurement unit 402 (see FIG. 7). The processor 3001 executes, for example, calculation processing related to control signals transmitted and received between the device mechanism section 455, the sample preparation section 440, and the specimen suction mechanism 450. The processor 3001 executes, for example, calculation processing related to transmission and reception of information between the computer 310X. The computer 310X has, for example, a function of displaying an analysis result transmitted from the analysis unit 303X based on the processing of the processor 3001. The computer 310X transmits, for example, a measurement order to the analysis unit 303X. The measurement order is, for example, input by a user via an input device of the computer 310X. The measurement order is, for example, transmitted from a host computer to the computer 310X. The processor 3001 executes, for example, processing related to reading program data from the storage section 3004, loading the program into the main memory 3017, and transmitting and receiving data between the main memory 3017. The above-mentioned processes executed by the processor 3001 are required to be executed in a predetermined order, for example. For example, if the processes required to control the device mechanism section 455, the sample preparation section 440, and the specimen aspiration mechanism 450 are A, B, and C, they may be required to be executed in the order of B, A, and C. Since the processor 3001 often executes successive processes that depend on such an order, increasing the number of arithmetic units (which may be called "processor cores", "cores", etc.) does not necessarily increase the processing speed.

[0208] On the other hand, the parallel processing processor 3002 executes a large amount of routine calculation processing, such as, for example, calculation of matrix data including a large number of elements. In this embodiment, the parallel processing processor 3002 executes parallel processing in which at least a part of the processing for analyzing waveform data according to the deep learning algorithm 60 is parallelized. The deep learning algorithm 60 includes, for example, a large amount of matrix operations. The deep learning algorithm 60 may include, for example, at least 100 matrix operations, or may include at least 1000 matrix operations. The parallel processing processor 3002 has a plurality of arithmetic units, and each of these arithmetic units can execute a matrix operation simultaneously. That is, the parallel processing processor 3002 can execute a matrix operation by each of the plurality of arithmetic units in parallel as a parallel process. For example, the matrix operation included in the deep learning algorithm 60 can be divided into a plurality of arithmetic processes that are not order-dependent with each other. The arithmetic processes divided in this way can be executed in parallel by each of the plurality of arithmetic units. These arithmetic units are sometimes called "processor cores", "cores", etc.

[0209] By executing such parallel processing, it is possible to speed up the calculation processing of the measurement unit 402 as a whole. Processing such as matrix calculation included in the deep learning algorithm 60 may be called, for example, "Single Instruction Multiple Data" (SIMD). The parallel processing processor 3002 is suitable for such SIMD calculation, for example. Such a parallel processing processor 3002 may be called a vector processor.

[0210] As described above, the processor 3001 is suitable for executing various and complicated processes. On the other hand, the parallel processing processor 3002 is suitable for executing a large amount of standardized processes in parallel. By executing a large amount of standardized processes in parallel, the TAT (Turn Around Time) required for the calculation process is shortened. Note that the parallel processing performed by the parallel processing processor 3002 is not limited to matrix operations. For example, when the parallel processing processor 3002 executes a learning process according to the deep learning algorithm 50, a differential operation related to the learning process, etc., may be the target of the parallel processing.

[0211] The number of arithmetic units of the processor 3001 is, for example, a dual core (number of cores: 2), a quad core (number of cores: 4), or an octa core (number of cores: 8). On the other hand, the parallel processing processor 3002 has, for example, at least 10 arithmetic units (number of cores: 10) and can execute 10 matrix operations in parallel. Some parallel processing processors 3002 have, for example, several tens of arithmetic units. Some parallel processing processors 3002 have, for example, at least 100 arithmetic units (number of cores: 100) and can execute 100 matrix operations in parallel. Some parallel processing processors 3002 have, for example, at least 1000 arithmetic units (number of cores: 1000) and can execute 1000 matrix operations in parallel. Some parallel processing processors 3002 have, for example, several thousand arithmetic units.

[0212] FIG. 48 shows a configuration example of the parallel processing processor 3002. The parallel processing processor 3002 includes a plurality of arithmetic units 3200 and a RAM 3201. Each of the arithmetic units 3200 executes arithmetic processing of matrix data in parallel. The RAM 3201 stores data related to the arithmetic processing executed by the arithmetic units 3200. The RAM 3201 is a memory having a capacity of at least 1 gigabyte. The RAM 3201 may be a memory having a capacity of 2 gigabytes, 4 gigabytes, 6 gigabytes, 8 gigabytes, or 10 gigabytes or more. The arithmetic unit 3200 obtains data from the RAM 3201 and executes arithmetic processing. The arithmetic unit 3200 may be called a "processor core", a "core", or the like.

[0213] 49, 50, and 51 show an overview of the arithmetic processing executed by the parallel processing processor 3002 based on the control of the analysis software 3100 operating on the processor 3001. FIG. 49 shows an example of the configuration of the parallel processing processor 3002 that executes the arithmetic processing. The parallel processing processor 3002 has a plurality of arithmetic units 3200 and a RAM 3201. The processor 3001 that executes the analysis software 3100 instructs the parallel processing processor 3002 to execute at least a part of the arithmetic processing required when analyzing the waveform data by the deep learning algorithm 60. The processor 3001 instructs the parallel processing processor 3002 to execute the arithmetic processing related to the analysis of the waveform data based on the deep learning algorithm 60. All or at least a part of the waveform data corresponding to the signal detected by the FCM detection unit 460 (see FIG. 4) is stored in the main memory 3017. The data stored in the main memory 3017 is transferred to the RAM 3201 of the parallel processing processor 3002. The data stored in the main memory 3017 is transferred to the RAM 3201 by, for example, a direct memory access (DMA) method. Each of the multiple arithmetic units 3200 of the parallel processing processor 3002 executes arithmetic processing on the data stored in the RAM 3201 in parallel. Each of the multiple arithmetic units 3200 obtains necessary data from the RAM 3201 and executes arithmetic processing. Data corresponding to the arithmetic results is stored in the RAM 3201 of the parallel processing processor 3002. The data corresponding to the arithmetic results is transferred from the RAM 3201 to the main memory 3017 by, for example, a DMA method.

[0214] FIG. 50 shows an overview of the matrix operation executed by the parallel processing processor 3002. When analyzing the waveform data according to the deep learning algorithm 60, a matrix multiplication (matrix operation) is performed. The parallel processing processor 3002 executes, for example, a plurality of arithmetic processes related to the matrix operation in parallel. (a) of FIG. 50 shows a formula for the matrix multiplication. In the formula shown in (a), a matrix c is obtained by multiplying a matrix a with n rows and n columns by a matrix b with n rows and n columns. As exemplified in FIG. 50, the formula is written in a multi-layer loop syntax. (b) of FIG. 50 shows an example of arithmetic processes executed in parallel by the parallel processing processor 3002. The formula exemplified in FIG. 50(a) can be divided into n×n arithmetic processes, which is the number of combinations of a loop variable i in the first layer and a loop variable j in the second layer. Each of the arithmetic processes divided in this way is an arithmetic process that does not depend on each other, and therefore can be executed in parallel.

[0215] Fig. 51 is a conceptual diagram showing that the multiple arithmetic processes illustrated in Fig. 50(b) are executed in parallel by the parallel processor 3002. As shown in Fig. 51, each of the multiple arithmetic processes is assigned to one of the multiple arithmetic units 3200 included in the parallel processor 3002. Each of the arithmetic units 3200 executes the assigned arithmetic processes in parallel with each other. In other words, each of the arithmetic units 3200 executes the divided arithmetic processes simultaneously.

[0216] 51, for example, information regarding the probability that a cell corresponding to the waveform data belongs to each of a plurality of cell types is obtained. Based on the results of the calculations, the processor 3001 executing the analysis software 3100 performs an analysis regarding the cell type of the cell corresponding to the waveform data. The results of the calculations are stored in the RAM 3201 of the parallel processor 3002, and transferred from the RAM 3201 to the main memory 3017. The processor 3001 obtains the results of the analysis based on the results of the calculations stored in the main memory 3017, and stores them in the storage unit 3004.

[0217] The calculation of the probability that a component in a sample belongs to each of the multiple classification types may be performed by a processor other than the parallel processing processor 3002. For example, the calculation results by the parallel processing processor 3002 may be transferred from the RAM 3201 to the main memory 3017, and the processor 3001 may calculate information regarding the probability that a component corresponding to each waveform data belongs to each of the multiple classification types based on the calculation results read from the main memory 3017. Furthermore, the calculation results by the parallel processing processor 3002 may be transferred from the RAM 3201 to the analysis unit 303X (see FIG. 47), and a processor mounted on the analysis unit 303X may calculate information regarding the probability that a component corresponding to each waveform data belongs to each of the multiple classification types.

[0218] <How to classify white blood cells> 1. First fluorescent dye and second fluorescent dye Next, an example of the first fluorescent dye and the second fluorescent dye will be described. In this example, an example in which two types of fluorescent dyes, the first fluorescent dye and the second fluorescent dye, are used to stain white blood cells in a sample will be described. In particular, a combination of the first fluorescent dye and the second fluorescent dye that is suitable for accurately counting basophils even in a blood sample where some time has passed since blood collection will be described below.

[0219] The second fluorescent dye is a fluorescent dye having a maximum absorption in a wavelength range different from that of the first fluorescent dye. That is, the second fluorescent dye is a fluorescent dye that emits fluorescence at a wavelength that can be distinguished from the fluorescence from the first fluorescent dye. The first fluorescent dye and the second fluorescent dye can each be appropriately selected from known fluorescent dyes that have the property of binding to nucleic acids in blood cells such as white blood cells.

[0220] The first and second fluorescent dyes are excited by light irradiated from a light source included in the flow cytometer. The light may be irradiated from one light source or from two light sources. When light is irradiated from one light source, the light irradiated from the light source may be light capable of exciting both the first and second fluorescent dyes. Such light is preferably light having a plurality of wavelengths, for example, white light. Alternatively, when the first and second fluorescent dyes have maximum absorption in a wavelength range close to the extent that they can be excited by light of one wavelength, the light irradiated from the light source may be light of that one wavelength. For example, when one of the first and second fluorescent dyes has maximum absorption in a wavelength range of 400 to 520 nm and the other has maximum absorption in a wavelength range of 300 to 420 nm, both fluorescent dyes can be excited by light having a central wavelength of 400 to 420 nm, for example, light of 455 nm. Alternatively, for example, if the maximum absorption of the first fluorescent dye and the second fluorescent dye is in the wavelength range of 630 to 660 nm, and one of the first fluorescent dye or the second fluorescent dye emits fluorescence having a peak in the wavelength range of 660 to 670 nm, and the other emits fluorescence having a peak in a wavelength range longer than 670 nm, both fluorescent dyes can be excited by light having a central wavelength in the range of 630 to 655 nm, for example, 633 nm, and the light generated from each fluorescent dye can be detected separately.

[0221] When light is emitted from two light sources, the emitted light may be two types of light, that is, light of a first wavelength capable of exciting a first fluorescent dye, and light of a second wavelength capable of exciting a second fluorescent dye. The second wavelength is different from the first wavelength. The wavelength can be appropriately determined depending on the type of fluorescent dye. For example, the first wavelength is 315 to 490 nm, preferably 400 to 450 nm, and more preferably 400 to 410 nm. The second wavelength is 610 to 750 nm, preferably 620 to 700 nm, and more preferably 633 to 643 nm.

[0222] When two excitation lights are used, the first fluorescent dye is preferably a dye that has a maximum absorption in the wavelength range of 400 to 490 nm, is excited by absorbing light in that wavelength range and emits fluorescence, and has the property of binding to nucleic acids (especially DNA) in blood cells. For example, fluorescent dyes having an acridine skeleton, 4',6-diamidino-2-phenylindole dihydrochloride (DAPI), Hoechst series Hoechst3342, Hoechst33258, Hoechst334580, etc. can be mentioned.

[0223] Examples of fluorescent dyes having an acridine skeleton include proflavine, 9-aminoacridine, acridine orange, acridine yellow G, acriflavine, basic yellow 9, ethacridine lactate, eucrisine GGNX, proflavine hemisulfate, 3,6-bis(dimethylamino)acridine (Rhoduline Orangen), and 3,6-diamino-2,7,10-trimethyl-acridinium chloride. Among these, acridine yellow G is preferred. Alternatively, the second fluorescent dye may be a commercially available fluorescent dye.

[0224] When two excitation lights are used, the second fluorescent dye is preferably a dye that has a maximum absorption in the wavelength range of 610 to 750 nm, is excited by absorbing light in that wavelength range and emits fluorescence, and has the property of binding to nucleic acids (especially RNA) in blood cells. For example, propidium iodide, ethidium bromide, ethidium-acridine heterodimer, ethidium diazide, ethidium homodimer-1, ethidium homodimer-2, ethidium monoazide, trimethylenebis[[3-[[4-[[(3-methylbenzothiazol-3-ium)-2-yl]methylene]-1,4-dihydroquinolin]-1-yl]propyl]dimethylaminium] tetraiodide (TOTO-1), 4-[(3-methylbenzothiazol-2(3H)-ylidene)methyl]-1-[3-(trimethylaminio)propyl]quinolinium diiodide (TO -PRO-1), N,N,N',N'-tetramethyl-N,N'-bis[3-[4-[3-[(3-methylbenzothiazol-3-ium)-2-yl]-2-propenylidene]-1,4-dihydroquinolin-1-yl]propyl]-1,3-propanediaminium tetraiodide (TOTO-3) or 2-[3-[[1-[3-(trimethylaminio)propyl]-1,4-dihydroquinolin]-4-ylidene]-1-propenyl]-3-methylbenzothiazol-3-ium diiodide (TOPRO-3), fluorescent dyes represented by the following formula (V), and combinations thereof.

[0225] [ka]

[0226] In formula (V), R 1 and R 4 R is a hydrogen atom, a methyl group, an ethyl group, or an alkyl group having 6 to 18 carbon atoms, and when one of them is an alkyl group having 6 to 18 carbon atoms, the other is a hydrogen atom, a methyl group, or an ethyl group. 2 and R 3are the same or different and are a methyl group, an ethyl group, a methoxy group, or an ethoxy group. Z is a carbon atom having a sulfur atom, an oxygen atom, or a methyl group. n is 0, 1, 2, or 3. X - is an anion.

[0227] In formula (V), the alkyl group having 6 to 18 carbon atoms may be either linear or branched. Among the alkyl groups having 6 to 18 carbon atoms, alkyl groups having 6, 8 or 10 carbon atoms are preferred.

[0228] In formula (V), R 1 and R 4 Examples of the substituent on the benzyl group include an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, or an alkynyl group having 2 to 20 carbon atoms. Among these, a methyl group or an ethyl group is particularly preferred.

[0229] In formula (V), R 2 and R 3 Examples of the alkenyl group include alkenyl groups having 2 to 20 carbon atoms. 2 and R 3 The alkoxy group includes an alkoxy group having a carbon number of 1 to 20. Among them, a methoxy group or an ethoxy group is particularly preferable.

[0230] In formula (V), the anion X - As, F - , Cl - , Br - and I - Halogen ions such as CF3SO3 - , BF4 - , ClO4 - etc.

[0231] As the fluorescent dye represented by the above formula (V), a fluorescent dye represented by the following formula is preferred.

[0232] [ka]

[0233] Commercially available staining reagents containing the above-mentioned second fluorescent dye alone may also be used, such as Fluorocell WDF (Sysmex Corporation) and Stromatolyser 4DS (Sysmex Corporation).

[0234] When one excitation light is used, the preferred combination of the first fluorescent dye and the second fluorescent dye may be different. For example, when 633 nm light is used as the excitation light, the maximum absorption of the first fluorescent dye and the second fluorescent dye may be in the wavelength range of 630 to 660 nm, the second fluorescent dye may emit fluorescence having a peak in the wavelength range of 660 to 670 nm, and the first fluorescent dye may emit fluorescence having a peak in the wavelength range longer than 670 nm. As such a combination, for example, DRAQ5, DRAQ7 or DRAQ9 (BioStatus) may be used as the first fluorescent dye, and the fluorescent dye represented by the above formula V may be used as the second fluorescent dye.

[0235] The first fluorescent dye and the second fluorescent dye are preferably used in the form of a solution. The solvent is not particularly limited as long as it can dissolve each of the above fluorescent dyes. Examples include water, organic solvents, and mixtures thereof. The organic solvent is preferably a solvent that can be mixed with water, and examples of the organic solvent include alcohols having 1 to 6 carbon atoms, ethylene glycol, diethylene glycol, polyethylene glycol, DMSO, and the like.

[0236] As described with reference to Figures 2 and 5, the first fluorescent dye and / or the second fluorescent dye may be contained in a single reagent container 200, or alternatively, as described with reference to Figure 6, they may be separately contained in different reagent containers 200A, 200B.

[0237] 2. Hemolysis Reagent

[0238] The above-mentioned first and second fluorescent dyes are used in combination with a hemolysis reagent, particularly preferably a hemolysis reagent containing a surfactant. The surfactant can hemolyze red blood cells in a specimen and damage the cell membranes of white blood cells to an extent that the first and second fluorescent dyes can penetrate the cell membranes. Examples of surfactants include nonionic surfactants, cationic surfactants, and combinations thereof. The hemolysis reagent preferably contains a nonionic surfactant.

[0239] An example of the nonionic surfactant is one represented by the following formula (I): R 1 -R 2 -(CH2CH2O) n -H (I) (In the formula, R 1 is an alkyl group, an alkenyl group, or an alkynyl group having 8 to 25 carbon atoms; R 2 represents an oxygen atom, -(COO)- or the following formula (II):

[0240] [ka] and n is equal to or greater than 23 and equal to or less than 25, or equal to 30.

[0241] In formula (I), n is preferably 23 or 25, more preferably 23. When n is 23 or more and 25 or less, the concentration of the nonionic surfactant represented by formula (I) in the hemolysis reagent is 1700 ppm or more, preferably 1750 ppm or more. When n is 23 or more and 25 or less, the concentration of the nonionic surfactant represented by formula (I) in the measurement sample is 2300 ppm or less, preferably 2200 ppm or less.

[0242] When n is 30, the concentration of the nonionic surfactant represented by formula (I) in the hemolysis reagent is 1900 ppm or more, preferably 2000 ppm or more, more preferably 2100 ppm or more. When n is 30, the concentration of the nonionic surfactant represented by formula (I) in the measurement sample is 2300 ppm or less, preferably 2200 ppm.

[0243] Specific examples of the nonionic surfactant represented by formula (I) include polyoxyethylene alkyl ether, polyoxyethylene sterol, polyoxyethylene castor oil, polyoxyethylene sorbitol fatty acid ester, polyoxyethylene alkylamine, polyoxyethylene polyoxypropylene alkyl ether, and combinations thereof. Among them, polyoxyethylene alkyl ether is preferred. The polyoxyethylene alkyl ether is preferably polyoxyethylene (23) cetyl ether, polyoxyethylene (25) cetyl ether, polyoxyethylene (30) cetyl ether, and at least one selected from the group consisting of polyoxyethylene (23) cetyl ether, polyoxyethylene (25) cetyl ether, and combinations thereof, and further preferably polyoxyethylene (23) cetyl ether. The nonionic surfactant contained in the hemolysis reagent may be one type or two or more types. The hemolysis reagent may further contain a nonionic surfactant other than the nonionic surfactant represented by formula (I).

[0244] The hemolysis reagent may further contain a cationic surfactant. Examples of the cationic surfactant include quaternary ammonium salt surfactants, pyridium salt surfactants, and combinations thereof. Examples of the quaternary ammonium salt surfactants include surfactants represented by the following formula (III) having a total carbon number of 9 to 30. The cationic surfactant contained in the hemolysis reagent may be one type, or two or more types.

[0245] [ka]

[0246] In formula (III), R 1 is an alkyl or alkenyl group having 6 to 18 carbon atoms; R 2 and R 3 are the same or different and each is an alkyl group or an alkenyl group having 1 to 4 carbon atoms; R 4 is an alkyl group or an alkenyl group having 1 to 4 carbon atoms, or a benzyl group; X - is a halogen ion.

[0247] In formula (III), R 1 is preferably an alkyl or alkenyl group having 6, 8, 10, 12 or 14 carbon atoms, and is particularly preferably a linear alkyl group. 1 Examples of R include an octyl group, a decyl group, and a dodecyl group. 2 and R 3 are preferably the same or different and each is a methyl group, an ethyl group, or a propyl group. 4 is preferably a methyl group, an ethyl group, or a propyl group.

[0248] An example of the pyridinium salt surfactant is a surfactant represented by the formula (IV).

[0249] [ka]

[0250] In formula (IV), R 1 is an alkyl or alkenyl group having 6 to 18 carbon atoms; X - is a halogen ion.

[0251] In formula (IV), R 1 is preferably an alkyl or alkenyl group having 6, 8, 10, 12 or 14 carbon atoms, and is particularly preferably a linear alkyl group. 1Examples of the alkyl group include octyl, decyl and dodecyl groups.

[0252] The concentration of the cationic surfactant in the hemolysis reagent can be appropriately selected depending on the type of surfactant. The concentration of the cationic surfactant is 10 ppm or more. The concentration of the cationic surfactant is preferably 400 ppm or more, more preferably 500 ppm or more, and even more preferably 600 ppm or more. The concentration of the cationic surfactant is 10000 ppm or less. The concentration of the cationic surfactant is preferably 1000 ppm or less, more preferably 800 ppm or less, and even more preferably 700 ppm or less.

[0253] The hemolysis reagent may contain a buffer substance to keep the pH constant. Examples of the buffer include inorganic acid salts, organic acid salts, Good's buffers, and combinations thereof. Examples of the inorganic acid salts include phosphate, borate, and combinations thereof. Examples of the organic acid salts include citrate, malate, and combinations thereof. Examples of the Good's buffer include MES, Bis-Tris, ADA, PIPES, Bis-Tris-Propane, ACES, MOPS, MOPSO, BES, TES, HEPES, HEPPS, Tricine, Tris, Bicine, TAPS, and combinations thereof.

[0254] The hemolysis reagent may further contain an aromatic organic acid. In this specification, aromatic organic acid means an acid having at least one aromatic ring in the molecule and its salt. Examples of aromatic organic acids include aromatic carboxylic acids and aromatic sulfonic acids. Examples of aromatic carboxylic acids include phthalic acid, benzoic acid, salicylic acid, hippuric acid, salts thereof, and combinations thereof. Examples of aromatic sulfonic acids include p-aminobenzenesulfonic acid, benzenesulfonic acid, salts thereof, and combinations thereof. The aromatic organic acid contained in the hemolysis reagent may be one type or two or more types. In addition, aromatic organic acids may exhibit a buffering effect. When an aromatic organic acid exhibiting a buffering effect is used, the addition of a buffering agent is optional and may be combined with the above-mentioned buffering agent.

[0255] When the hemolysis reagent contains an aromatic organic acid, the concentration of the aromatic organic acid is not particularly limited, but is preferably 20 mM or more, more preferably 25 mM or more, from the viewpoint of the ability to distinguish monocytes and lymphocytes. The concentration of the aromatic organic acid contained in the hemolysis reagent is preferably 50 mM or less, more preferably 45 mM or less.

[0256] The hemolysis reagent is preferably a liquid reagent. The solvent is not particularly limited as long as it can dissolve each component such as the surfactant. Examples of the solvent include water, organic solvents, and mixtures thereof. Examples of the organic solvent include solvents that are miscible with water, such as alcohols having 1 to 6 carbon atoms, ethylene glycol, diethylene glycol, polyethylene glycol, and dimethyl sulfoxide (DMSO).

[0257] The pH of the hemolysis reagent is not particularly limited, but the pH is preferably 5.5 or higher. More preferably, the pH is 5.7 or higher, and even more preferably, the pH is 5.9 or higher. The pH is preferably 7.2 or lower. More preferably, the pH is 6.9 or lower, and even more preferably, the pH is 6.6 or lower. The pH can be adjusted using a known base (such as sodium hydroxide) or acid (such as hydrochloric acid).

[0258] In the hemolysis reagent, the osmotic pressure is not particularly limited, but from the viewpoint of the hemolysis efficiency of red blood cells, it is preferably 150 mOsm / kg or less, more preferably 130 mOsm / kg or less, and most preferably 110 mOsm / kg or less. A suitable osmotic pressure regulator may be added to adjust the osmotic pressure. Examples of the osmotic pressure regulator include sugars, amino acids, organic solvents, sodium chloride, and combinations thereof.

[0259] The hemolytic reagent may be a commercially available hemolytic reagent for measuring blood cells, such as Lyzacell WDF (Sysmex Corporation) or Lyzacell WDFII (Sysmex Corporation).

[0260] The hemolysis reagent is contained in, for example, a reagent container R1 (see FIG. 5), and is delivered to the chamber 420 by the method described above and mixed with the blood sample.

[0261] 3.White blood cell classification method Next, as a sixth embodiment of the present invention, a method for classifying white blood cells into subpopulations using the above-mentioned first and second fluorescent dyes will be described. This method is carried out, for example, by the steps shown in the flowchart of Fig. 52. Each step will be described below. Fig. 52 is an example of a flowchart of a method for classifying white blood cells into subpopulations, and Fig. 53 is a flowchart of a preferred embodiment for realizing the method of Fig. 52.

[0262] [Step S0: Preparing a measurement sample] In this step, a measurement sample is prepared by mixing a specimen containing white blood cells, a hemolysis reagent containing a surfactant, a first fluorescent dye, and a second fluorescent dye. The hemolysis reagent, the first fluorescent dye, and the second fluorescent dye are preferably those described above.

[0263] The specimen may be any sample that contains white blood cells or may contain blood cells, and is not limited to whole blood. Blood cells refer to cells known to be contained in whole blood, such as white blood cells, red blood cells, and platelets. Samples that contain white blood cells or may contain blood cells are body fluid samples collected from mammals, preferably humans. Examples of specimens include whole blood, ascites, synovial fluid, pleural effusion, cerebrospinal fluid, bone marrow fluid, bronchoalveolar lavage fluid, peritoneal lavage fluid, urine, and samples collected by apheresis, etc.

[0264] The first and second fluorescent dyes are mixed with the specimen and hemolysis reagent so that their concentrations (final concentrations) in the measurement sample are within a predetermined range. The upper limit of the preferred final concentration of the first fluorescent dye in the measurement sample is 1000 ppm or less, preferably 100 ppm or less, more preferably 10 ppm or less. The lower limit of the preferred final concentration of the first fluorescent dye in the measurement sample is 0.001 ppm or more, preferably 0.01 ppm or more, more preferably 0.1 ppm or more. The upper limit of the preferred final concentration of the second fluorescent dye in the measurement sample is 1000 ppm or less, preferably 100 ppm or less, more preferably 10 ppm or less. The lower limit of the preferred final concentration of the second fluorescent dye in the measurement sample is 0.001 ppm or more, preferably 0.01 ppm or more, more preferably 0.1 ppm or more.

[0265] The mixing ratio of the hemolysis reagent, the staining reagent, and the specimen is preferably 1000:1 or more:1 or more, for example, expressed as a volume ratio. More preferably, it is 1000:10 or more:10 or more, and even more preferably, it is 1000:15 or more:15 or more. Furthermore, the mixing ratio of the hemolysis reagent, the staining reagent, and the specimen is preferably 1000:50 or less:50 or less, for example, expressed as a volume ratio. More preferably, it is 1000:30 or less:30 or less, and even more preferably, it is 1000:25 or less:25 or less. The mixing ratio of the fluorescent dye and the specimen may be the same or different.

[0266] [Step S1: Detecting optical information] In this step, light is irradiated onto particles in the prepared measurement sample, and optical information including first fluorescence information based on the fluorescence from the first fluorescent dye, second fluorescence information based on the fluorescence from the second fluorescent dye, and scattered light information is detected. The optical information is preferably detected by the FCM detection unit 460 (see FIG. 2). "Particles in the measurement sample" refers to granular objects contained in the measurement sample that can be individually measured by the FCM detection unit 460. Specifically, the measurement sample is first introduced into the flow cell 4113 of the FCM detection unit 460, and light is irradiated onto each particle in the measurement sample as it passes through the flow cell. Then, the scattered light and fluorescence emitted from the particle are measured to detect the optical information. The particles in the measurement sample may include not only cells such as white blood cells, but also non-cellular particles such as remnants of hemolyzed red blood cells (red blood cell ghosts), platelet aggregates, and lipid particles.

[0267] The optical information detected is scattered light information and fluorescence information. Since this method uses two types of fluorescent dyes, fluorescence information corresponding to the fluorescence emitted from each fluorescent dye is detected. That is, first fluorescence information based on the fluorescence from the first fluorescent dye and second fluorescence information based on the fluorescence from the second fluorescent dye are detected. When light is irradiated from one light source, scattered light information detected by the irradiation of the light is detected as scattered light information. When light of a first wavelength and light of a second wavelength are irradiated from two light sources, respectively, first scattered light information detected by the irradiation of light of the first wavelength and second scattered light information detected by the irradiation of light of the second wavelength are detected as scattered light information.

[0268] Examples of scattered light include forward scattered light (e.g., scattered light with a light receiving angle of 0 degrees to about 20 degrees) and side scattered light (e.g., scattered light with a light receiving angle of about 20 degrees to about 90 degrees). Examples of scattered light information and fluorescent information include the peak values ​​(heights of the pulse peaks) of scattered light and fluorescent light, pulse area, pulse width, transmittance, Stokes shift, ratio, changes over time, and values ​​correlated therewith. The forward scattered light information is not particularly limited as long as it is information that reflects the size of the cell. The side scattered light information is not particularly limited as long as it is information that reflects internal information such as the complexity of the cell structure, granular characteristics, nuclear structure, and degree of lobulation. Examples of scattered light information include forward scattered light peak value and side scattered light peak value, and more preferably side scattered light peak height. Examples of fluorescent information include fluorescent peak height.

[0269] The first scattered light information is preferably the side scattered light peak value detected by irradiation with light of a first wavelength (hereinafter also referred to as "first side scattered light intensity"). The second scattered light information is preferably the side scattered light peak value detected by irradiation with light of a second wavelength (hereinafter also referred to as "second side scattered light intensity"). The first fluorescent information is preferably the fluorescent peak value of the first fluorescent dye (hereinafter also referred to as "first fluorescent intensity"). The second fluorescent information is preferably the fluorescent peak value of the second fluorescent dye (hereinafter also referred to as "second fluorescent intensity").

[0270] For example, an FCM capable of detecting first fluorescence information based on the fluorescence from a first fluorescent dye, second fluorescence information based on the fluorescence from a second fluorescent dye, and scattered light information can be used for detecting optical information. As such an FCM detection unit, for example, an FCM equipped with an optical system and detection system shown in Figs. 7 and 8 can be used.

[0271] [Step S2: Step of selecting a cell population containing basophils] In this step, a cell population including basophils is selected from particles in a measurement sample based on optical information including the first fluorescence information. It is believed that this step can remove populations that affect the fractionation of basophils in advance. As described above, in a specimen that has been collected for some time, populations that affect the fractionation of basophils may occur. The present inventor has found that there is a distinguishable difference in the first fluorescence information between such populations and normal basophils. For example, as shown in FIG. 56(c) of Reference Example 1 described later, each subpopulation of normal white blood cells usually has approximately the same fluorescence intensity in a scattergram with the first fluorescence intensity (blue-purple) and the second side scattered light intensity (red) as the two axes. Here, the dots on the scattergram represent individual particles measured by FCM. The side scattered light may be the first side scattered light (blue-purple). On the other hand, as shown in Fig. 57(a) to Fig. 57(c) of Reference Example 2, in a specimen collected some time ago, a population with a lower first fluorescence intensity than the subpopulation of normal white blood cells appears. This population is also found in the region where normal basophils appear on a scattergram with the second fluorescence intensity and the side scattered light intensity as the two axes, and therefore interferes with the fractionation of basophils. Therefore, by selecting a cell population containing basophils based on optical information including the first fluorescence information, it becomes possible to remove a population that affects the fractionation of basophils from the measured particles. In the sixth embodiment, the cell population containing basophils is preferably a population of white blood cells containing basophils.

[0272] When normal white blood cells are stained with a hemolysis reagent and a first fluorescent dye, each subpopulation of the white blood cells has approximately the same first fluorescent intensity, as described above. Therefore, a cell population containing basophils can be selected by the first fluorescent information. Specifically, when the optical information containing the first fluorescent information is the first fluorescent intensity, a particle population having a first fluorescent intensity greater than a predetermined threshold is selected from particles in a measurement sample. In this case, a cell population containing lymphocytes, monocytes, neutrophils, eosinophils, and basophils is considered to be selected as a cell population containing basophils.

[0273] Alternatively, as shown in steps S2-1 and S2-2 of the flow chart in FIG. 53, a scattergram based on the first fluorescent information and the scattered light information (also referred to as a first scattergram) may be created, and a cell population containing basophils may be selected based on the scattergram. The scattered light information may be the first scattered light information. For example, when the optical information including the first fluorescent information is the first fluorescent intensity and the side scattered light intensity, a particle population that appears in a predetermined region where the first fluorescent intensity is greater than a predetermined threshold value in a scattergram having the first fluorescent intensity and the side scattered light intensity as two axes can be selected. Either the first side scattered light intensity or the second side scattered light intensity may be used as the side scattered light intensity. For example, as shown in FIG. 56(c), in a scattergram with the first fluorescent intensity on the vertical axis and the second side scattered light intensity on the horizontal axis, each subpopulation of white blood cells is distributed in a substantially horizontal line. That is, the region in which each subpopulation appears in the scattergram is also known. The predetermined region where the first fluorescent intensity is greater than a predetermined threshold value can be preset as a region where a subpopulation of white blood cells including basophils appears. Such a region can be, for example, a region where five subpopulations of lymphocytes, monocytes, neutrophils, eosinophils, and basophils appear, a region where four subpopulations of lymphocytes, monocytes, neutrophils, and basophils appear, or a region where three subpopulations of lymphocytes, monocytes, and basophils appear. It is considered that a cell population of white blood cells including basophils is selected by selecting a group of particles that appears in such a region by gating.

[0274] The predetermined threshold value corresponding to the first fluorescence intensity is not particularly limited and can be determined appropriately. For example, blood known to contain basophils or blood that has been collected for some time may be measured in advance by FCM using the above-mentioned hemolysis reagent and the first fluorescent dye to obtain the first fluorescence intensity for leukocytes, and the obtained value may be used as the predetermined threshold value. In addition, the threshold value and the region where the cell population containing basophils appears may be set in advance by accumulating data on the region where each subpopulation of leukocytes appears from the first scattergram created based on such measurement.

[0275] [Step S3: Classifying leukocytes contained in a cell population into subpopulations] In this step, white blood cells contained in a cell population containing basophils are classified into subpopulations based on the second fluorescent information and the scattered light information. The scattered light information may be second scattered light information. Since populations that affect the fractionation of basophils are removed by the above-mentioned step of selecting cell populations, it is considered that white blood cells contained in the selected cell population can be classified into each subpopulation containing basophils with high accuracy. In this step, for example, as shown in S3-1 and S3-2 of the flowchart in FIG. 53, a scattergram (also called a second scattergram) based on the second fluorescent information and the scattered light information is created, and white blood cells contained in a cell population containing basophils may be classified into subpopulations based on the scattergram. The scattered light information may be second scattered light information. Specifically, a scattergram with the second fluorescent intensity and the side scattered light intensity as the two axes is created. The side scattered light intensity may be the second side scattered light intensity. When the selected cell population is a population of white blood cells including basophils, i.e., a cell population including lymphocytes, monocytes, neutrophils, eosinophils, and basophils, in the scattergram, each subpopulation appears in a different region and can be classified into five groups, for example, as shown in FIG. 56(a). Since the cell size, amount of nucleic acid, internal structure, etc. differ from one another for each subpopulation, the second fluorescent intensity and side scattered light intensity detected for each subpopulation differ, resulting in the classification in this manner. Thus, the selected cell population can be classified into a lymphocyte population, a monocyte population, a neutrophil population, an eosinophil population, and a basophil population.

[0276] Alternatively, when the selected cell population is a cell population containing lymphocytes, monocytes, neutrophils, and basophils, in a scattergram with the second fluorescent intensity and the side scattered light intensity as the two axes, each subpopulation appears in a different region and can be classified into four subpopulations. That is, the selected cell population can be classified into a lymphocyte population, a monocyte population, a neutrophil population, and a basophil population.

[0277] Alternatively, when the selected cell population is a cell population containing lymphocytes, monocytes, and basophils, in a scattergram with the second fluorescent light intensity and the side scattered light intensity as the two axes, each subpopulation appears in a different region and can be classified into three subpopulations, as shown in Figure 59(b) of Example 1. That is, the selected cell population can be classified into a lymphocyte population, a monocyte population, and a basophil population.

[0278] [Step S4: Counting cells classified into the basophil group] In this step, cells classified into the basophil group are counted among the subpopulations of leukocytes classified as described above. In the step of selecting the cell population containing basophils, groups that affect the fractionation of basophils are removed, so it is considered that the cells classified into the basophil group do not substantially contain cells other than basophils (e.g., degraded neutrophils and lymphocytes). Therefore, the counting step allows the basophils in the measurement sample to be counted more accurately. The counting is preferably performed by analyzing the second scattergram with an appropriate analysis software to obtain the number of cells classified into the basophil group.

[0279] [Seventh embodiment] The method of classifying white blood cells into subpopulations according to the seventh embodiment is carried out, for example, by the steps shown in the flow chart of FIG. 54. The preparation of the measurement sample (step S10) and the detection of optical information (step S11) can be carried out in the same manner as in the sixth embodiment. In the seventh embodiment, between the step of detecting optical information (step S11) and the step of selecting a cell population containing basophils (steps S14 and S15), a step of selecting a cell population containing nucleated cells from particles in the measurement sample based on the first fluorescence information (steps S12 and S13) is carried out. The first fluorescence information is preferably the first fluorescence intensity. Specifically, a histogram is created for the particles in the measurement sample, with the number of particles on the vertical axis and the first fluorescence intensity on the horizontal axis (step S12). Since the first fluorescent dye has the property of binding to the nucleic acid of cells as described above, when the particles are nucleated cells, the first fluorescence intensity is higher than that of particles not containing nucleic acid. Using this, particles whose first fluorescent intensity is equal to or greater than a predetermined threshold or within a predetermined range in the histogram are selected as nucleated cells (step S13). By such selection, particles without nuclei, such as red blood cell ghosts, can be removed from the particles in the measurement sample. The predetermined threshold and the lower limit of the predetermined range here may be values ​​that are sufficient to remove particles with a very low first fluorescent intensity, such as red blood cell ghosts. In the seventh embodiment, a cell population containing basophils may be selected from a cell population containing nucleated cells. The process after the step of selecting a cell population containing basophils can be performed in the same manner as in the sixth embodiment.

[0280] [Eighth embodiment] The method of classifying white blood cells into subpopulations in the eighth embodiment is carried out, for example, by the steps shown in the flow chart of FIG. 55. The preparation of the measurement sample (step S20) and the detection of optical information (step S21) can be carried out in the same manner as in the sixth embodiment. The selection of the cell population containing nucleated cells (steps S22, S23) can be carried out in the same manner as in the seventh embodiment. It is possible to arbitrarily decide whether or not to select the cell population containing nucleated cells. In the eighth embodiment, between the step of detecting optical information (step S21) and the step of selecting the cell population containing basophils (steps S27, S28), a step of classifying the particle or cell population containing nucleated cells in the measurement sample into four subpopulations, a lymphocyte population, a monocyte population, an eosinophil population, and a population containing both neutrophils and basophils, based on the second fluorescent information and the scattered light information (steps S24, S25). This classification itself can be carried out in the same manner as the step of classifying white blood cells contained in the cell population into subpopulations in the sixth embodiment. Specifically, a second scattergram is created based on the second fluorescent light information and the scattered light information (step S24 ), and based on the scattergram, the white blood cells contained in the cell population containing particles or nucleated cells in the measurement sample are classified into subpopulations (step S25). If time has passed since the collection of the specimen, the white blood cells in the specimen are poorly fractionated, and in particular, the fractionation of basophils and neutrophils becomes difficult. If the measured specimen is a specimen with such poor fractionation, in the classification process, the cell population containing particles or nucleated cells in the measurement sample is classified into four subpopulations: a lymphocyte population, a monocyte population, an eosinophil population, and a population containing both neutrophils and basophils. For example, the particles classified into a population containing both neutrophils and basophils are counted. If the measured specimen is a specimen with good fractionation, in the classification process, the cell population containing particles or nucleated cells in the measurement sample may be classified into five subpopulations: a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil population, and a basophil population (step S26).

[0281] In the eighth embodiment, after classifying the cell population containing particles or nucleated cells in the measurement sample into four subpopulations, similarly to the sixth embodiment, the steps of selecting the cell population containing basophils (steps S27 and S28), classifying the leukocytes contained in the cell population into subpopulations (step S29), and counting the cells classified into the basophil population (step S30) are performed. Then, based on the results obtained by the step of classifying into four subpopulations and the result obtained by the step of counting the cells classified into the basophil population, a step (step S31) of classifying the particles in the measurement sample into five subpopulations, namely, a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil population, and a basophil population, is further performed. For example, the number of particles classified into the neutrophil population can be obtained by subtracting the number of particles obtained by the step of counting the cells classified into the basophil population (step S30) from the number of particles of the population containing both neutrophils and basophils obtained by the step of classifying into four subpopulations (step S25). As a result, in the eighth embodiment, the particles in the measurement sample can be classified into five subpopulations, namely, a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil population, and a basophil population. If the particles in the measurement sample can be classified into five subpopulations of white blood cells at the stage of the process of classifying into four subpopulations, the classification result can be compared with the classification result finally obtained in the eighth embodiment.

[0282] The method for classifying leukocytes according to the present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. EXAMPLES

[0283] The specimen containing leukocytes, the hemolysis reagent, the staining dye, the analyzer, and the measurement method used in the following examples will be described.

[0284] [Sample containing white blood cells] (Whole blood sample) 5 mL of whole blood was collected from healthy volunteers in EDTA-2K blood collection tubes. After collection, the blood was stored at room temperature (25±2°C), and samples were taken from the blood collection tubes 4 hours, 24 hours, 42 hours, and 72 hours later to prepare the test samples.

[0285] (Pre- and post-basophil isolation samples) A 1 mL sample with high basophil count (>2%) was prepared, and 0.5 mL of it was used as the pre-basophil isolation sample. The remaining sample was used to remove red blood cells using HetaSep (SCT ST-07906) and recover white blood cells. Basophil cells were then recovered using Basophil Isolation Kit II, human (Miltenyi Biotec 130-092-662), and this was used as the post-basophil isolation sample.

[0286] [Hemolysis reagent] The Lizacell WDFII (Sysmex Corporation) was used.

[0287] [Staining reagent] (First fluorescent dye) Acridine Yellow G (Kanto Chemical Co., Ltd.) was used.

[0288] (Second fluorescent dye) As the second fluorescent dye, dye compound A described in U.S. Patent No. 6,004,816 was used. U.S. Patent No. 6,004,816 is incorporated herein by reference. Dye compound A is represented by the formula (V) above, 1 is a methyl group, and R 2 and R 3 is a hydrogen atom, and R 4 is n-octyl, n is 1, Z is a sulfur atom, and X - CF3SO3 - The compound had the following structural formula:

[0289] [ka]

[0290] As described in US Patent 6004816, dye compound A was obtained by the following process. 1 equivalent of 3-methyl-2-methylbenzothiazolium methanesulfate and 3 equivalents of N,N-diphenylformamidine were heated and stirred in acetic acid on a 90°C oil bath for 1.5 hours. The reaction solution was poured into hexane, and the red oily product was further suspended and washed in hexane to remove the acetic acid. The crude product was recrystallized from ethyl acetate-hexane (yield 48%). 1 equivalent of 1-octyl lepidinium triflate and pyridine were added to this, and the mixture was heated and stirred on a 90°C oil bath for 3 hours. The reaction solution was concentrated, and the remaining blue crude product was purified by flash chromatography with methanol-chloroform to obtain dye compound A as a deep dark blue powder (yield 62%). The results of physical property tests (TLC, 1H-NMR, MASS, etc.) of this deep dark blue powder dye compound A are described in US Patent 6004816. The maximum absorption spectrum of dye compound A was 629 nm. Ta.

[0291] The dye compound A (27.5 mg) and acridine yellow G (25 mg) were dissolved in special grade ethylene glycol (1 L) to prepare a staining reagent.

[0292] [Analyzer] A modified multi-item automated blood cell analyzer XN-1000 (Sysmex Corporation) was created. The XN-1000 was equipped with a semiconductor laser light source that emits red light (633 nm), a detector for red forward scattered light, a detector for red side scattered light, and a detector for receiving fluorescence excited by red light (light of 660 nm or more). The XN-1000 was modified to add a semiconductor laser light source that emits blue-violet light (405 nm), a detector for blue-violet scattered light, and a fluorescence detector that detects fluorescence excited by blue light (450 to 600 nm). In addition, the analyzer was designed to analyze measurement data detected regarding red and blue-violet scattered light information and fluorescence information generated by the first and second fluorescent dyes, and to display the desired scattergram. Measurements were performed using the analyzer thus created.

[0293] [Measurement method] The measurement samples were prepared and measured according to the manual attached to the XN-1000, except that the staining reagent containing the first and second fluorescent dyes prepared as described above was used instead of Fluorocell WDF (Sysmex Corporation), which is a staining reagent for the XN-1000 (Sysmex Corporation). Data analysis was performed using FCS reanalysis software (Flowjo). The prepared measurement sample contained 1000 μL of Lysacell WDFII as a hemolysis reagent, 17 μL of whole blood, and 20 μL of staining reagent. The dilution ratio of the staining reagent at this time was 51.85, and the final concentrations of the first and second fluorescent dyes in the measurement sample were 0.53 ppm and 0.48 ppm, respectively.

[0294] Reference Example 1: Location of basophils Measurements were performed on the above-mentioned pre-basophil separation specimen and post-basophil separation specimen. From the obtained optical information, a scattergram (first scattergram) was created with the horizontal axis representing the side scattered light intensity of the second wavelength (red wavelength) and the vertical axis representing the first wavelength (blue-violet fluorescence), and a scattergram (second scattergram) was created with the horizontal axis representing the side scattered light intensity of the second wavelength (red wavelength) and the vertical axis representing the second wavelength (red fluorescence).

[0295] The obtained scattergrams are shown in Figures 56(a) to 56(d). In the second scattergram (Figure 56(a)) of the sample before basophil separation, the five types of white blood cell populations are distributed apart in the two axial directions, and it is understood that the fractionation is good. On the other hand, in the first scattergram (Figure 56(c)) of the sample before basophil separation, it is understood that the five white blood cell populations are distributed apart in the horizontal axis (side scattering) direction, but all appear in the region of almost the same fluorescence intensity in the vertical axis (blue-violet fluorescence) direction. In addition, in the measurement result of the sample after basophil separation, basophils appear in the region of lower fluorescence intensity than lymphocytes in the second scattergram (Figure 56(b)), and appear in the position of fluorescence intensity equal to that of lymphocytes and monocytes in the first scattergram (Figure 56(d)), and it was possible to confirm the fractionation state in the whole blood sample.

[0296] Reference Example 2: Cluster appearance positions that change over time Measurements were performed on whole blood samples taken 4 hours, 48 ​​hours, and 72 hours after collection to detect optical information. Then, a population included in the low value region of the vertical axis (blue-violet fluorescence) that increases with time in the first scattergram was selected (gated), and the population was plotted on the second scattergram.

[0297] The results of the first scattergram are shown in Figures 57(a) to 57(d), and the results of the second scattergram are shown in Figures 58(a) to 58(c). From Figures 58(a) to 58(c), it can be seen that the number of cells contained in the area where basophils appear increases with the passage of time after blood collection. It was also found that the cause of this increase was the cells gated in the first scattergram (which appeared in the area enclosed by a square in Figures 57(a) to 57(c)). That is, it was revealed that in the second scattergram, neutrophils shift toward the low scattered light value side with the passage of time after blood collection, and the group thought to be derived from neutrophils, which overlaps with the area where basophils appear, is the group that appears on the low blue-violet fluorescence value side in the first scattergram.

[0298] Example 1: Basophil count 1 (one-stage gating (no gating of nucleated cells by blue-violet fluorescence)) Measurements were performed on whole blood samples taken 4 hours, 24 hours, 48 ​​hours, and 72 hours after collection, and optical information was detected. Then, a first scattergram was created and a population included in a preset region (see FIG. 59(a)) where the blue-violet fluorescence was greater than a predetermined value was selected. The population appearing in this region includes lymphocytes, monocytes, and basophils, but does not include cells derived from neutrophils, which shift to the region where basophils appear over time after collection. The first scattergram for the sample taken 24 hours after collection is shown in FIG. 59(a).

[0299] Then, a second scattergram was created for the selected population. The population included in the previously set basophil appearance region in this second scattergram was counted. The second scattergram for the sample taken 24 hours after blood collection is shown in Figure 59(b).

[0300] Example 2: Basophil count 2 (two-stage gating (with gating of nucleated cells by blue-violet fluorescence) corresponding to the second embodiment) and Comparative Example 1 The following analysis was performed using the optical information obtained in Example 1. First, cells (nucleated cells) whose obtained blue-violet fluorescence exceeded a predetermined threshold were selected. A histogram of the sample taken 24 hours after blood collection at this time is shown in FIG. 60(a). After this, the same analysis as in Example 1 was performed to count basophils. The first and second scattergrams are shown in FIG. 60(b) and FIG. 60(c).

[0301] On the other hand, basophils were counted based on a second scattergram prepared from the red side scattered light intensity and the red fluorescent light intensity out of the acquired optical information (Comparative Example 1).

[0302] The basophil count values ​​obtained for samples collected at various times after blood collection by Example 2 and Comparative Example 1 are shown in Figure 61. As can be seen from Figure 61, by employing the method of the present invention, the increase in the basophil count value was suppressed compared to the Comparative Example, even for samples collected long ago.

[0303] Further, Fig. 62 shows the second scattergrams obtained by the methods of Example 2 and Comparative Example 1 for the samples taken 4 hours and 48 hours after blood collection. As is clear from the scattergrams, it can be seen that the increase in basophils is suppressed by the method of the present invention.

[0304] Example 3: Five-part classification of white blood cells Based on the optical information obtained in Example 1, the leukocytes were classified into five subpopulations. Among the optical information, the red side scattered light intensity and the red fluorescence were used to create a second scattergram, and the cells contained in each of the previously set lymphocyte appearance regions, monocyte appearance regions, eosinophil appearance regions, and neutrophil and basophil appearance regions were counted. That is, at this stage, the leukocytes were classified into four types of populations and counted.

[0305] Thereafter, a population including lymphocytes, monocytes, and basophils was selected in a first scattergram with the horizontal axis representing blue-violet side scattered light and the vertical axis representing blue-violet fluorescence, and the basophils in the selected population were counted in a second scattergram with the horizontal axis representing red side scattered light intensity and the vertical axis representing red fluorescence, in the same manner as in Example 1. The basophil count obtained here was subtracted from the count of the population of neutrophils and basophils out of the four types obtained previously, to count the neutrophil population, and the five subpopulations of leukocytes were counted together with the previous results.

[0306] The thus obtained counts of the five types of white blood cells for each time period after blood collection are shown in Table 2. Meanwhile, from the acquired optical information, a second scattergram was created using the red side scattered light intensity and red fluorescence, and the cells contained in the respective areas in which lymphocytes, monocytes, eosinophils, neutrophils, and basophils appeared, which had been set in advance, were counted (Comparative Example 2). In other words, the white blood cells were classified into five subpopulations by the second scattergram created initially. The obtained results are also shown in Table 2.

[0307] [Table 2]

[0308] As is clear from Table 2, according to the present invention, the increase in the number of basophils over time after blood collection is suppressed. In addition, it was found that a single measurement can simultaneously and accurately classify 5 types of leukocytes. [Explanation of symbols]

[0309] 54: chamber; 60, 442: reagent container holder; 60a, 60b, 60c, 60d, 60e: holder portion; 62: reagent container holding portion; 64: piercer (aspiration tube); 100: blood collection tube; 200, 200A, 200B, 300: reagent container; 300X, 301X, 302X, 303X: analyzer (analysis unit, analysis portion); 310X: computer; 400, 400X, 401, 402: measuring device (measurement unit); 410, 410a, 410b: reagent container; 420: chamber; 430, 430a, 430b: liquid delivery mechanism; 431, 431a, 431b: liquid delivery tube; 431x, 431ax, 431bx: first end; 431y, 431ay, 431by: second end, 432, 432a, 432b: liquid delivery section, 433: pump (quantitation section), 440, 440A, 440B, 440X, 441: sample preparation section, 450: specimen suction mechanism, 451: specimen suction nozzle, 452: suction and discharge mechanism (pump), 460, 460x: FCM detection section (detection section), 4111: light source, 4111a: first light source, 4111b: second light source, 4113: flow cell, 4116: forward scattered light receiving element, 4118, 4118a, 4118b, 4118c: dichroic mirror, 4121a, 4121b: side scattered light receiving element, 4122a, 4122b: side fluorescence light receiving element, C: cell (component), D1: first light receiving section, D2: second light receiving section, D3: third light receiving section, F1, F2: fluorescent dye, FC: flow cell, FSC: forward scattered light, SFL: side fluorescence, SSC: side scattered light

Claims

1. A measuring device for analyzing cells contained in a specimen, comprising: a chamber for preparing a measurement sample in which the cells are stained with first and second fluorescent dyes contained in a reagent supplied from at least one reagent container; A first light source that irradiates light of a first wavelength; a second light source that irradiates light having a second wavelength that is longer than the first wavelength; a detection unit that acquires, in response to irradiation of the measurement sample flowing through a flow cell with light, a first side scattered light signal corresponding to side scattered light by the first light source and a first fluorescent light signal corresponding to fluorescence emitted from the cells stained with the first fluorescent dye, and a second side scattered light signal corresponding to side scattered light by the second light source, a forward scattered light signal corresponding to forward scattered light, and a second fluorescent light signal corresponding to fluorescence emitted from the cells stained with the second fluorescent dye; an analysis unit that analyzes the cells based on the first side scattered light signal, the first fluorescent light signal, the second side scattered light signal, the forward scattered light signal, and the second fluorescent light signal; A measuring device comprising:

2. The measurement device according to claim 1 , wherein the detection section is configured to acquire a forward scattered light signal corresponding to the second fluorescent dye out of the first and second fluorescent dyes.

3. The measuring device according to claim 1 or 2, wherein the detection section includes a forward scattered light receiving element that receives only forward scattered light corresponding to the light emitted from the second light source.

4. The detection unit is a first side scattered light receiving element disposed to receive side scattered light corresponding to light from the first light source; a first fluorescence receiving element disposed to receive fluorescence corresponding to light from the first light source; a second side scattered light receiving element disposed to receive side scattered light corresponding to the light from the second light source; a forward scattered light receiving element disposed to receive forward scattered light corresponding to the light from the second light source without receiving forward scattered light corresponding to the light from the first light source; The measuring device according to claim 1 or 2, further comprising: a second fluorescence receiving element arranged to receive fluorescence corresponding to the light from the second light source.

5. the first wavelength is between 400 nm and 450 nm; The measuring device according to any one of claims 1 to 4, wherein the second wavelength is 620 nm to 700 nm.

6. the first light source emits light of the first wavelength, which is blue-violet; The measurement device according to any one of claims 1 to 5, wherein the second light source emits light of the second wavelength, which is red.

7. a liquid delivery unit for delivering the reagent from the reagent container to the chamber through a liquid delivery tube provided between the reagent container and the chamber; The measuring device according to any one of claims 1 to 6, wherein the liquid delivery section delivers the reagent from the reagent container to the chamber via a liquid delivery tube having a first end disposed in the reagent container and a second end connected to the chamber.

8. The measuring device according to claim 7 , wherein the first end of the liquid delivery tube is fixed at a predetermined position within the reagent container.

9. 9. The measurement device according to claim 8, wherein the first end of the liquid delivery tube is fixed at the predetermined position while a plurality of measurement samples respectively corresponding to a plurality of specimens are prepared.

10. a mechanism for inserting the first end of the liquid delivery tube into the reagent container and disposing the first end at a predetermined position within the reagent container; The measuring device according to any one of claims 7 to 9.

11. 11. The measuring device according to claim 7, wherein the liquid delivery section further comprises a metering section for delivering a fixed amount of the reagent in the reagent container to the chamber via the liquid delivery tube.

12. The measuring device according to any one of claims 1 to 11, wherein the reagent container contains 20 mL or more and 100 mL or less of the reagent.

13. 13. The measuring device according to claim 1, wherein a storage period of the reagent contained in the reagent container is 60 days or more and 90 days or less at a temperature in an environment in which the measuring device is installed.

14. 14. The measuring device according to claim 13, wherein the storage period of the reagent contained in the reagent container is from 60 to 90 days at a temperature of 15°C to 30°C.

15. The measuring device according to any one of claims 1 to 14, wherein the reagent container has a bag-shaped reagent storage portion for storing the reagent.

16. The measuring device according to claim 15 , wherein the reagent container contains 200 mL or more and 500 mL or less of the reagent.

17. 17. The measuring device according to claim 15, wherein the storage period of the reagent contained in the reagent container is from 75 days to one year at a temperature in an environment in which the measuring device is installed.

18. 18. The measuring device according to claim 17, wherein the storage period of the reagent contained in the reagent container is from 75 days to one year at a temperature of 15°C to 30°C.

19. The measuring device according to any one of claims 1 to 18, wherein the cells are stained with a first compound constituting the first fluorescent dye and a second compound constituting the second fluorescent dye.

20. The measuring device according to any one of claims 1 to 19, wherein the reagent does not contain an antibody.

21. The measurement device according to any one of claims 1 to 20, wherein the measurement sample is prepared by staining the cells in the specimen with the first and second fluorescent dyes mixed with a hemolytic agent that damages the cell membrane to allow the first and second fluorescent dyes to permeate the cell membrane.

22. The measuring device according to any one of claims 1 to 21, wherein the analysis unit analyzes the cells based on classification of the cells using the first side scattered light signal and the first fluorescent light signal, and classification of the cells using the second side scattered light signal, the forward scattered light signal, and the second fluorescent light signal.

23. The measuring device according to any one of claims 1 to 22, wherein the analysis unit analyzes the first side scattered light signal, the first fluorescent light signal, the second side scattered light signal, the forward scattered light signal and the second fluorescent light signal using an artificial intelligence algorithm.

24. The measurement device according to claim 23 , wherein the analysis unit executes the matrix calculation by the artificial intelligence algorithm in parallel processing by a parallel processing processor.

25. 25. The measuring device according to claim 23 or 24, wherein the artificial intelligence algorithm is a deep learning algorithm.

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