Blood cell analyzers and blood cell analysis methods

Abstract

This application relates to a blood cell analyzer and a blood cell analysis method. Specifically, a portion of the aspirated blood sample, a diluent, and a fluorescent staining agent are mixed in a first reaction chamber to prepare a first test sample, and first optical information is obtained from the particles in the first test sample after being irradiated with light. Another portion of the blood sample is mixed with a hemolysin in a second reaction chamber to prepare a second test sample, and colorimetric optical information is obtained from the second test sample. Based on the first optical information, reticulocyte, platelet, red blood cell, and first white blood cell counts of the blood sample are obtained, the first white blood cell count including at least the percentage of lymphocytes, monocytes, and granulocytes. The hemoglobin concentration of the blood sample is determined based on the colorimetric optical information. This application can reduce the reagent costs required to obtain the test results of the blood sample.

Description

Technical Field

[0001] This application relates to the field of blood analysis, and in particular to blood cell analyzers and methods for blood cell analysis. Background Technology

[0002] A complete blood count (CBC) is a basic clinical examination. The results generally include white blood cell count, red blood cell count, platelet count, hemoglobin concentration, and reticulocyte count. It also includes scatter plots or histograms of white blood cells, red blood cells, platelets, and reticulocytes obtained during the test to help doctors make clinical diagnoses of patients.

[0003] Existing blood cell analyzers use flow cytometry for optical measurement of white blood cells and fluorescence staining for optical measurement of reticulocytes (or platelets), while colorimetric methods are used to measure hemoglobin concentration. In optical measurement of white blood cells, the count and classification of white blood cells, including the count and classification of neutrophils, lymphocytes, monocytes, eosinophils, and basophils, are obtained. In optical measurement of reticulocytes, parameters such as red blood cell count, platelet count, and reticulocyte count are obtained.

[0004] The principle of colorimetric methods is that different concentrations of the test sample solution result in different colors. Generally, the higher the concentration, i.e., the higher the hemoglobin content, the darker the solution color; conversely, the lower the concentration, i.e., the lower the hemoglobin content, the lighter the solution color. The depth of the test sample solution color directly affects its light absorption; the darker the solution, the more light it absorbs and the less light it transmits, and vice versa. Therefore, by measuring the intensity of light transmitted through the test sample, the concentration of the solution can be determined, and thus the hemoglobin concentration of the test sample can be identified.

[0005] However, in existing blood cell analyzers, it is usually necessary to prepare three different test samples and perform three tests to obtain white blood cell test results, reticulocyte test results, and hemoglobin concentration, resulting in high reagent costs and large blood volumes. Summary of the Invention

[0006] In order to at least partially solve the above-mentioned technical problems, the objective of this application is to provide a blood cell analyzer and a blood cell analysis method, which can simultaneously obtain white blood cell detection results, reticulocyte detection results, and hemoglobin concentration while reducing reagent costs and blood usage.

[0007] To achieve the above-mentioned objectives of this application, a first aspect of this application provides a blood cell analyzer, comprising:

[0008] A sampling device is used to collect blood samples for testing.

[0009] A sample preparation apparatus is used to mix a portion of the blood sample to be tested, a diluent, and a fluorescent staining agent in a first reaction cell to prepare a first test sample, and to mix another portion of the blood sample to be tested and a hemolysin in a second reaction cell to prepare a second test sample;

[0010] An optical detection device includes a liquid driving unit, a flow chamber, a light source, and a photodetector. The liquid driving unit is used to drive the first test sample from the first reaction cell through the flow chamber. The light source is used to illuminate the first test sample passing through the flow chamber. The photodetector is used to detect the first optical information generated by the first test sample after being illuminated by light when passing through the flow chamber.

[0011] A colorimetric detection device is used to detect the second test sample in the second reaction cell to obtain colorimetric optical information;

[0012] The data processing device is configured as follows:

[0013] Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained. The first white blood cell detection result includes at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes, wherein the granulocytes include neutrophils and eosinophils.

[0014] The hemoglobin concentration of the blood sample to be tested is obtained based on the colorimetric optical information.

[0015] A second aspect of this application provides a method for blood cell analysis, comprising:

[0016] Collect the blood sample to be tested;

[0017] A portion of the blood sample to be tested, a diluent, and a fluorescent staining agent are mixed in a first reaction cell to prepare a first test sample. Particles in the first test sample in the first reaction cell are then passed one by one through an optically irradiated detection area to obtain the first optical information generated by the particles in the first test sample after being irradiated by light.

[0018] Another portion of the blood sample to be tested and a hemolysin are mixed in a second reaction chamber to prepare a second test sample, and the second test sample in the second reaction chamber is detected by colorimetry to obtain colorimetric optical information.

[0019] Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained. The first white blood cell detection result includes at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes, wherein the granulocytes include neutrophils and eosinophils.

[0020] The hemoglobin concentration of the blood sample to be tested is determined based on the colorimetric optical information.

[0021] In the technical solutions proposed in this application, only a first test sample for identifying platelets, reticulocytes, and erythrocytes and a second test sample for measuring hemoglobin concentration need to be prepared to obtain the reticulocyte detection results, platelet detection results, erythrocyte detection results, leukocyte detection results, and hemoglobin concentration of the blood sample to be tested, thereby reducing reagent costs and blood usage. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the structure of a blood cell analyzer according to some embodiments of this application.

[0023] Figure 2 This is a schematic diagram of the structure of an optical detection device according to some embodiments of this application.

[0024] Figure 3 This is a schematic diagram of the structure of a colorimetric detection apparatus according to some embodiments of this application.

[0025] Figure 4 The image shows the light signal generated when particles in a first measurement sample according to some embodiments of this application are irradiated with light.

[0026] Figure 5 A first scatter plot of FS-FL generated based on first optical information according to some embodiments of this application.

[0027] Figure 6 A second scatter plot of FS-SS generated based on first optical information according to some embodiments of this application.

[0028] Figure 7 A second scatter plot of FS-SS generated from first optical information according to other embodiments of this application.

[0029] Figure 8 A third scatter plot of FS-SS generated from second optical information according to some embodiments of this application.

[0030] Figure 9 This is a schematic flowchart of a blood cell analysis method according to some embodiments of this application. Detailed Implementation

[0031] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0032] It should be noted that the terms "first, second, and third" used in the embodiments of this application are merely to distinguish similar objects and do not represent a specific order of objects. It can be understood that "first, second, and third" can be interchanged in a specific order or sequence where permitted.

[0033] It will be understood by those skilled in the art that, unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0034] To facilitate subsequent explanations, a brief explanation of some terms used below will be provided first:

[0035] 1) Scatter plot: A two-dimensional or three-dimensional graph generated by a blood cell analyzer, displaying two-dimensional or three-dimensional feature information of multiple particles. The X, Y, and Z axes of a scatter plot each represent a characteristic of each particle. For example, in a scatter plot, the X axis represents the intensity of forward scattered light, the Y axis represents the fluorescence intensity, and the Z axis represents the intensity of side scattered light.

[0036] 2) Cell clusters: Clusters of particles with the same characteristics distributed in a certain area of ​​a scatter plot, such as white blood cell clusters, as well as neutrophil clusters, lymphocyte clusters, monocyte clusters, eosinophil clusters, or basophil clusters within white blood cells.

[0037] 3) Blood shadow: Fragment particles obtained by dissolving red blood cells and platelets in blood with a hemolytic reagent.

[0038] Currently, blood cell analyzers can test human blood and blood samples from other mammals (such as dogs, cats, and horses), birds, and fish. Typically, blood cell analyzers use the DIFF channel (white blood cell differential channel) to count and classify white blood cells, classifying them into five types: lymphocytes (Lym), monocytes (Mon), neutrophils (Neu), eosinophils (Eos), and eosinophils (Eos). In addition, blood cell analyzers use the RET channel (reticulocyte detection channel, or platelet optical detection channel) to obtain reticulocyte counts, red blood cell counts, and platelet counts.

[0039] The blood cell analyzer used in this application uses flow cytometry technology combining laser scattering and fluorescence staining to classify and count particles in a sample. For example, the detection principle of reticulocytes, platelets, and erythrocytes in the blood cell analyzer is as follows: First, a blood sample is drawn and treated with diluent and fluorescent dye. Then, particles in the blood sample pass one by one through a detection aperture irradiated by a laser beam. When the laser beam irradiates the particles, the characteristics of the particles themselves (such as volume, staining degree, size and content of cell contents, nuclear density, etc.) can block or change the direction of the laser beam, thereby generating scattered light at various angles corresponding to their characteristics. This scattered light is received by a signal detector to obtain relevant information about the particle structure and composition. Forward scatter (FS) reflects the number and volume of particles, side scatter (SS) reflects the complexity of the internal cell structure (such as intracellular particles or the cell nucleus), and fluorescence (FL) reflects the content of nucleic acid substances in the cell. This optical information can be used to classify and count particles in the sample.

[0040] As mentioned in the background art, in order to identify leukocytes and reticulocytes in the blood and measure hemoglobin concentration while reducing reagent costs and blood volume, this application proposes a technical solution that can simultaneously obtain leukocyte detection results, reticulocyte detection results and hemoglobin concentration by preparing only two test samples.

[0041] Figure 1 This is a schematic diagram of a hematology analyzer according to some embodiments of this application. The hematology analyzer 100 includes a sample aspiration device 110, a sample preparation device 120, an optical detection device 130, a data processing device 140, and a colorimetric detection device (not shown). The hematology analyzer 100 also has a fluid system (not shown) for connecting the sample aspiration device 110, the sample preparation device 120, the optical detection device 130, and the colorimetric detection device to facilitate fluid transfer between these devices.

[0042] The sampling device 110 is used to draw up the blood sample to be tested.

[0043] The sample preparation apparatus 120 is used to mix a portion of the blood sample to be tested, a diluent, and a fluorescent staining agent in a first reaction cell to prepare a first test sample, and to mix another portion of the blood sample to be tested and a hemolysin in a second reaction cell to prepare a second test sample.

[0044] In this embodiment, the diluent may include, for example, tris(hydroxymethyl)aminomethane buffer. By adjusting the amount of tris(hydroxymethyl)aminomethane buffer, the osmotic pressure of the diluent is adjusted to approximately 200 mOsm / kg (the osmotic pressure of mammalian blood cells is generally around 260–320 mOsm / kg). When blood cells are mixed with the diluent, red blood cells and white blood cells absorb water in the hypotonic solution, resulting in an increase in volume, while the volume of platelets remains relatively unchanged.

[0045] In the embodiments of this application, the fluorescent staining agent is a fluorescent dye used to distinguish reticulocytes, erythrocytes, platelets, and leukocytes. For example, the function of the fluorescent staining agent is to stain RNA and DNA in the cell nucleus.

[0046] Because mature red blood cells are anucleate, their staining effect is minimal, while reticulocytes contain filamentous structures. Therefore, staining with fluorescent dyes increases the fluorescence intensity of reticulocytes, distinguishing them from mature red blood cells. Furthermore, since the RNA and DNA content in the nucleus of leukocytes is significantly higher than that in reticulocytes, leukocytes exhibit the strongest signal intensity in the fluorescence direction after staining with fluorescent dyes. Thus, by controlling the staining time, platelets, mature red blood cells, reticulocytes, and leukocytes can be clearly distinguished.

[0047] In the embodiments of this application, the hemolytic agent is used to dissolve red blood cells in the blood, breaking them into fragments, while maintaining the morphology of white blood cells essentially unchanged. For example, the hemolytic agent can be any one or a combination of several of cationic surfactants, nonionic surfactants, anionic surfactants, and amphiphilic surfactants; or, for example, the hemolytic agent can include at least one of alkyl glycosides, triterpenoid saponins, and steroidal saponins.

[0048] The optical detection device 130 includes a liquid driving unit, a flow chamber, a light source, and a photodetector. The liquid driving unit is used to drive the first test sample from the first reaction cell through the flow chamber. The light source is used to illuminate the first test sample passing through the flow chamber. The photodetector is used to detect the first optical information generated by the first test sample after being illuminated by light when passing through the flow chamber.

[0049] As can be understood here, the detection channel (also known as the RET channel) used to identify reticulocytes, platelets and red blood cells refers to the detection of the first test sample prepared by the sample preparation device 120 by the optical detection device 130.

[0050] The colorimetric detection device is used to detect the second test sample in the second reaction cell to obtain colorimetric optical information.

[0051] As can be understood, red blood cells in the blood rupture under the action of a hemolytic agent, releasing hemoglobin, which can then be measured by colorimetry to determine the hemoglobin concentration (HGB). The detection channel used to determine the hemoglobin concentration (also known as the HGB channel) refers to the detection of the second test sample prepared by the sample preparation device 120 by the colorimetric detection device.

[0052] The data processing device 140 is configured as follows:

[0053] Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained. The first white blood cell detection result includes at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes, wherein the granulocytes include neutrophils and eosinophils.

[0054] The hemoglobin concentration of the blood sample to be tested is obtained based on the colorimetric optical information.

[0055] In the aforementioned blood cell analyzer 100, only a first test sample for identifying platelets, reticulocytes, and erythrocytes and a second test sample for measuring hemoglobin concentration need to be prepared to obtain the reticulocyte detection results, platelet detection results, erythrocyte detection results, white blood cell detection results, and hemoglobin concentration of the blood sample to be tested, thereby reducing reagent costs and blood usage.

[0056] The blood cell analyzer 100 will be further described below with reference to some embodiments.

[0057] In some embodiments, the sampling device 110 has a sampling needle (not shown) for drawing up a blood sample to be tested. Furthermore, the sampling device 110 may also include, for example, a driving device for driving the sampling needle to quantitatively draw up the blood sample through the tip of the sampling needle. The sampling device 110 can deliver the collected blood sample to the sample preparation device 120.

[0058] In some embodiments, the sample preparation apparatus 120 may include a first reaction cell, a second reaction cell, and a reagent supply device (not shown). Each reaction cell is used to receive the blood sample to be tested drawn by the sampling device 110, and the reagent supply device provides processing reagents (including hemolysin, fluorescent staining agent, diluent, etc.) to the corresponding reaction cell, so that the blood sample to be tested drawn by the sampling device 110 is mixed with the processing reagents provided by the reagent supply device in the corresponding reaction cell to prepare a test sample (including a first test sample and a second test sample).

[0059] For example, the reagent supply device may include a first reagent supply unit and a second reagent supply unit. The sampling device 110 is used to partially distribute the collected blood sample to be tested into a first reaction chamber and a second reaction chamber, respectively. The first reagent supply unit is used to provide diluent and fluorescent staining agent to the first reaction chamber, thereby mixing and reacting the portion of the blood sample allocated to the first reaction chamber with the diluent and fluorescent staining agent to prepare a first test sample. The second reagent supply unit is used to provide a hemolysin to the second reaction chamber, thereby mixing and reacting the portion of the blood sample allocated to the second reaction chamber with the hemolysin to prepare a second test sample.

[0060] In some embodiments, a flow chamber refers to a chamber with a focused liquid flow suitable for detecting light scattering and fluorescence signals. When a particle, such as a blood cell, passes through the detection aperture of the flow chamber, the particle scatters an incident light beam from a light source, directed through the aperture, in various directions. Photodetectors can be positioned at one or more different angles relative to the incident light beam to detect the light scattered by the particle, thereby obtaining a light scattering signal. Since different particles have different light scattering characteristics, the light scattering signal can be used to distinguish different groups of particles.

[0061] Specifically, the light scattering signal detected near the incident light beam is generally referred to as the forward light scattering signal or the small-angle light scattering signal. In some embodiments, the forward light scattering signal can be detected at an angle of about 1° to about 10° with respect to the incident light beam. In other embodiments, the forward light scattering signal can be detected at an angle of about 2° to about 6° with respect to the incident light beam. The light scattering signal detected at about 90° with respect to the incident light beam is generally referred to as the side light scattering signal. In some embodiments, the side light scattering signal can be detected at an angle of about 65° to about 115° with respect to the incident light beam. Typically, the fluorescent signal emitted from blood cells stained with fluorescent dye is also generally detected at about 90° with respect to the incident light beam.

[0062] In some embodiments, the photodetector may include a forward-scattering light detector for detecting forward-scattering light signals, a side-scattering light detector for detecting side-scattering light signals, and a fluorescence detector for detecting fluorescence signals. Accordingly, the first optical information may include the forward-scattering light signal, the side-scattering light signal, and the fluorescence signal of particles in the first measurement sample after being irradiated by light.

[0063] Figure 2 A specific example of an optical detection device 130 is shown. This optical detection device 130 has a light source 101, a beam shaping assembly 102, a flow chamber 103, and a forward scattering detector 104 arranged sequentially in a straight line. A dichroic mirror 106 is arranged at a 45° angle to the line on one side of the flow chamber 103. A portion of the side light emitted by particles in the flow chamber 103 passes through the dichroic mirror 106 and is captured by a fluorescence detector 105 arranged at a 45° angle behind the dichroic mirror 106; the other portion of the side light is reflected by the dichroic mirror 106 and captured by a side scattering detector 107 arranged at a 45° angle in front of the dichroic mirror 106.

[0064] In some embodiments, such as Figure 3 As shown, the colorimetric detection apparatus may include a light source and a photometer. The light source is used to illuminate the second test sample in the second reaction cell, and the photometer is used to detect information about the light transmitted through the second test sample in the second reaction cell.

[0065] In some embodiments, the data processing device 140 is used to process and perform calculations on data to obtain the desired results. For example, it can generate two-dimensional or three-dimensional scatter plots based on various collected optical signals, and perform particle analysis on the scatter plots using a gating method. The data processing device 140 can also visualize intermediate or final calculation results and then display them through the display device 150. In this embodiment, the data processing device 140 is configured to implement the method steps, which will be described in further detail below.

[0066] In some embodiments, the data processing device 140 includes, but is not limited to, devices such as a central processing unit (CPU), a microcontroller unit (MCU), a field-programmable gate array (FPGA), and a digital signal data processing device (DSP) used to interpret computer instructions and process data in computer software. For example, the data processing device is used to execute various computer applications in a computer-readable storage medium, thereby enabling the blood cell analyzer 100 to perform corresponding detection procedures and analyze the optical information or optical signals detected by the optical detection device 130 in real time.

[0067] Furthermore, the hematology analyzer 100 may also include a first housing 160 and a second housing 170. The display device 150 may be, for example, a user interface. An optical detection device 130 and a data processing device 140 are disposed inside the second housing 170. A sample preparation device 120 is disposed, for example, inside the first housing 160, and the display device 150 is disposed, for example, on the outer surface of the first housing 160 and is used to display the detection results of the hematology analyzer.

[0068] The steps to be performed by the processor 140 to obtain white blood cell detection results, reticulocyte detection results, and hemoglobin concentration are further explained below with reference to some embodiments.

[0069] In some embodiments, the data processing device 140 may be configured to, when acquiring the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested based on the first optical information:

[0070] A first scatter plot is generated based on at least the forward scattered light signal FS and the fluorescence signal FL in the first optical information, and the reticulocyte detection results, platelet detection results, and red blood cell detection results are obtained based on the first scatter plot; and

[0071] A second scatter plot is generated based on at least the forward scattered light signal FS and the side scattered light signal SS in the first optical information, and the first white blood cell detection result is obtained based on the second scatter plot.

[0072] like Figure 4As shown, the first test sample in the first reaction cell is passed through the flow chamber. When the particles in the first test sample pass through the flow chamber, they are irradiated by the light source (e.g., a laser beam), generating a forward-scattered light signal FS (low angle) and a side-scattered light signal SS (90-degree high angle signal). After red blood cells, platelets, reticulocytes, and white blood cells are stained with fluorescent dyes, the particles in the first test sample, upon irradiation by the laser beam, will generate an excitation light signal with a higher wavelength. This excitation light signal has the same receiving angle (90 degrees) as the side-scattered light signal SS. A beam splitter is used to separate the side-scattered light signal SS from the excitation light signal; the separated excitation light signal is called the fluorescence signal FL. The forward-scattered light signal FS reflects cell volume, the side-scattered light signal SS reflects the complexity of the cell's internal structure (such as intracellular granules or the cell nucleus), and the fluorescence signal FL reflects the content of nucleic acid substances in the cell (such as RNA / DNA content).

[0073] The first optical information may include the forward scattered light signal FS, the side scattered light signal SS, and the fluorescence signal FL of the particles in the first measurement sample after being irradiated by light.

[0074] For example, Figure 5 A first scatter plot is shown, generated based on at least the forward scattered light signal FS and the fluorescence signal FL from the first optical information. The direction of the forward scattered light signal FS corresponds to the cell size, and the direction of the fluorescence signal FL corresponds to the intensity of the fluorescent dye staining. In this first scatter plot, from left to right along the direction of the fluorescence signal FL, the cells are mature erythrocytes, low-fluorescence reticulocytes, medium-fluorescence reticulocytes, high-fluorescence reticulocytes, and leukocytes.

[0075] Based on this first scatter plot, reticulocyte, platelet, and erythrocyte detection results can be obtained. The reticulocyte detection results can include reticulocyte count and reticulocyte classification, which includes low-fluorescence reticulocytes, medium-fluorescence reticulocytes, and high-fluorescence reticulocytes. The platelet detection results can include platelet count. The erythrocyte detection results can include erythrocyte count.

[0076] In some embodiments, the parameter results shown in Table 1 can be obtained by combining the first scatter plot and the hemoglobin concentration of the test sample.

[0077] Table 1

[0078]

[0079] For example, Figure 6A second scatter plot is shown, generated based on at least the forward scattered light signal FS and the side scattered light signal SS from the first optical information. The first white blood cell detection result obtained based on this second scatter plot includes at least the percentage of lymphocytes (RET_Lym), monocytes (RET_Mon), and granulocytes (RET_Gran). It should be understood that the granulocytes include neutrophils and eosinophils.

[0080] Furthermore, the first white blood cell detection result obtained based on the second scatter plot may also include lymphocyte RET_Lym count, monocyte RET_Mon count, and granulocyte RET_Gran count.

[0081] In some embodiments, the sample preparation device 120 may be further configured to add a chemical staining agent during the preparation of the first test sample, so that the data processing device 140 can distinguish eosinophils among the granulocytes based on the first optical information, such that the first white blood cell detection result includes at least the percentage of lymphocytes (RET_Lym), monocytes (RET_Mon), eosinophils (RET_Eos), and neutrophils (RET_Neu). Figure 7 As shown.

[0082] Table 2 shows at least some of the parameter results from the first white blood cell detection results obtained based on the first optical information after the addition of chemical staining agents.

[0083] Table 2

[0084]

[0085] Thus, by simply adding a chemical staining agent during the preparation of the first test sample for measuring platelets, reticulocytes, and erythrocytes, white blood cells can be classified into four types—lymphocytes (Lym), monocytes (Mon), neutrophils (Neu), and eosinophils (Eos)—through the detection of the first test sample. This eliminates the need for additional test samples for white blood cell counting and classification. In other words, the RET channel can simultaneously obtain platelet, reticulocyte, erythrocyte, and four-part white blood cell differential results, reducing reagent costs and blood usage.

[0086] Optionally, such as Figure 7As shown, the data processing device 140 can be further configured to identify basophils (RET_Baso) in the first test sample based on the first optical information, such that the first white blood cell detection result also includes the percentage of basophils (RET_Baso). That is, the first white blood cell detection result includes the percentage of lymphocytes (RET_Lym), monocytes (RET_Mon), eosinophils (RET_Eos), neutrophils (RET_Neu), and basophils (RET_Baso).

[0087] Table 3 shows the results of various parameters obtained after identifying basophils in the first test sample based on the first optical information.

[0088] Table 3

[0089]

[0090] Thus, by simply adding a chemical staining agent during the preparation of the first test sample for platelet determination, white blood cells can be classified into five categories—lymphocytes (Lym), monocytes (Mon), neutrophils (Neu), eosinophils (Eos), and basophils (Baso)—through the detection of the first test sample. This eliminates the need for additional test samples for white blood cell counting and classification. In other words, the RET channel can simultaneously obtain platelet, reticulocyte, erythrocyte, and five-part white blood cell differential results, reducing reagent costs and blood usage.

[0091] In some embodiments, the sample preparation apparatus 120 may be further configured to add the chemical staining agent either by mixing it with the diluent and / or the fluorescent staining agent or by adding it separately when preparing the first assay sample.

[0092] For example, the sample preparation apparatus 120 can be further configured to add the chemical staining agent in a manner mixed with the diluent when preparing the first test sample. As another example, the sample preparation apparatus 120 can be further configured to add the chemical staining agent in a manner mixed with the fluorescent staining agent when preparing the first test sample. Yet another example, the sample preparation apparatus 120 can be further configured to add the chemical staining agent in a manner simultaneously mixed with the diluent and the fluorescent staining agent when preparing the first test sample. Still another example, the sample preparation apparatus 120 can be further configured to add the chemical staining agent separately to the first reaction cell when preparing the first test sample.

[0093] In some embodiments, the chemical dye may be selected from acidic organic pigments.

[0094] Optionally, the chemical staining agent is selected from the Acid Blue series;

[0095] Optionally, the chemical dye is selected from the group consisting of Direct Blue, Acid Green, Acid Yellow, Acid Orange, Methyl Red, Methyl Orange, Aniline Blue, Alizarin Yellow, Reactive Black, Sudan Black B, and Azo Black E.

[0096] In some embodiments, the liquid driving unit in the optical detection device 130 may be further used to drive at least a portion of the second measurement sample from the second reaction cell through the flow chamber, and the light source may be further used to irradiate the at least a portion of the second measurement sample passing through the flow chamber with light, and the photodetector may be further used to detect the second optical information generated after the at least a portion of the second measurement sample is irradiated with light when passing through the flow chamber.

[0097] Here, the data processing device 140 is further configured to acquire a second white blood cell detection result of the blood sample to be tested based on the second optical information, wherein the second white blood cell detection result includes at least the white blood cell count, basophil count and basophil percentage.

[0098] It is understood that the detection channel for detecting basophils (also known as the basophil detection channel) refers to the detection of at least a portion of the second test sample prepared by the sample preparation device 120 by the optical detection device 130.

[0099] Table 4 shows at least some of the parameter results in the second white blood cell detection results obtained based on the second optical information.

[0100] Table 4

[0101]

[0102] In this way, at least a portion of the prepared second test sample can be used directly to count and classify basophils to obtain accurate white blood cell count, basophil count and basophil percentage, thus eliminating the need to prepare additional test samples for identifying basophils. This reduces reagent costs and blood usage while still obtaining accurate white blood cell test results.

[0103] In some embodiments, when the data processing device 140 obtains the second white blood cell detection result of the blood sample to be tested based on the second optical information, it generates a third scatter plot based on the forward scattered light signal FS and the side scattered light signal SS in the second optical information, and obtains the second white blood cell detection result based on the third scatter plot.

[0104] For example, Figure 8A third scatter plot is shown, generated based on the forward scattered light signal FS and the side scattered light signal SS from the second optical information. Based on this third scatter plot, basophils (Baso) in the second test sample can be identified.

[0105] In some embodiments, a more accurate five-part differential white blood cell test result can be obtained by combining the first white blood cell test result obtained through the RET channel and the second white blood cell test result obtained through at least partial detection of the second assay sample. That is, accurate reticulocyte, platelet, red blood cell, white blood cell, and hemoglobin concentration results can be obtained by preparing only a first assay sample for identifying platelets, reticulocytes, and red blood cells and a second assay sample for measuring hemoglobin concentration.

[0106] In some embodiments, the liquid drive unit is configured to drive at least a portion of the second test sample from the second reaction cell through the flow chamber after the colorimetric detection device detects the second test sample in the second reaction cell to obtain the colorimetric optical information, so that the photodetector detects the second optical information. That is, basophil counting and classification can be performed after the hemoglobin concentration determination.

[0107] In this way, the second assay sample in the second reaction chamber can be reused for basophil counting and classification, thereby further reducing the reagent cost and blood volume required for the test.

[0108] In other embodiments, the liquid drive unit is configured to drive at least a portion of the second test sample from the second reaction cell through the flow chamber before the colorimetric detection device detects the second test sample in the second reaction cell to obtain the colorimetric optical information, so that the photodetector detects the second optical information generated after the at least a portion of the second test sample is irradiated by light as it passes through the flow chamber. That is, basophil counting and classification can be performed before the determination of hemoglobin concentration.

[0109] Thus, since the hemoglobin concentration of the second test sample remaining in the second reaction chamber is measured after basophil counting and classification, there is sufficient time for the blood sample and reagents in the second reaction chamber to react fully before the hemoglobin concentration is measured, which improves the accuracy of the subsequent hemoglobin concentration measurement and thus improves the accuracy of the obtained hemoglobin concentration.

[0110] Accordingly, such as Figure 9 As shown in the embodiments of this application, a blood cell analysis method 200 is also proposed, comprising:

[0111] S210, collect the blood sample to be tested;

[0112] S220, a portion of the blood sample to be tested, a diluent, and a fluorescent staining agent are mixed in a first reaction cell to prepare a first test sample, and the particles in the first test sample in the first reaction cell are passed one by one through an optical detection area irradiated by light to obtain the first optical information generated by the particles in the first test sample after being irradiated by light.

[0113] S230, another part of the blood sample to be tested and the hemolysin are mixed in the second reaction cell to prepare the second test sample, and the second test sample in the second reaction cell is detected by colorimetry to obtain colorimetric optical information;

[0114] S240, based on the first optical information, acquire the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested, wherein the first white blood cell detection results include at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes, and the granulocytes include neutrophils and eosinophils; and

[0115] S250, determine the hemoglobin concentration of the blood sample to be tested based on the second optical information.

[0116] In some embodiments, in step S220, a chemical staining agent is additionally added when preparing the first test sample, so that in step S240, eosinophils in the granulocytes can be distinguished based on the first optical information, and the first white blood cell detection result includes at least the percentage of lymphocytes, the percentage of monocytes, the percentage of eosinophils, and the percentage of neutrophils.

[0117] Optionally, in step S240, basophils in the first test sample are also identified based on the first optical information, so that the first white blood cell detection result also includes the percentage of basophils.

[0118] In some embodiments, in step S220, the chemical staining agent is added either by mixing it into the diluent and / or the fluorescent staining agent or by adding it alone when preparing the first test sample.

[0119] In some embodiments, the chemical dye is selected from acidic organic pigments.

[0120] Optionally, the chemical staining agent is selected from the Acid Blue series;

[0121] Optionally, the chemical dye is selected from the group consisting of Direct Blue, Acid Green, Acid Yellow, Acid Orange, Methyl Red, Methyl Orange, Aniline Blue, Alizarin Yellow, Reactive Black, Sudan Black B, and Azo Black E.

[0122] In some embodiments, step S240 may include: generating a first scatter plot based on at least the forward-scattered light signal and the fluorescence signal in the first optical information, and obtaining the reticulocyte detection result, platelet detection result and red blood cell detection result based on the first scatter plot; and generating a second scatter plot based on at least the forward-scattered light signal and the side-scattered light signal in the first optical information, and obtaining the first white blood cell detection result based on the second scatter plot.

[0123] In some embodiments, the blood cell analysis method 200 may further include the following steps:

[0124] The particles in at least a portion of the second test sample in the second reaction cell are passed one by one through an optically irradiated detection area to obtain second optical information generated by the particles in at least a portion of the second test sample after being irradiated by light; and

[0125] The second white blood cell detection result of the blood sample to be tested is obtained based on the second optical information. The second white blood cell detection result includes at least the white blood cell count, basophil count and basophil percentage.

[0126] In some embodiments, obtaining the second white blood cell detection result of the blood sample to be tested based on the second optical information may include: generating a third scatter plot based on the forward scattered light signal and the side scattered light signal in the second optical information, and obtaining the second white blood cell detection result based on the third scatter plot.

[0127] In some embodiments, before or after the second test sample in the second reaction cell is detected by colorimetry to obtain the colorimetric optical information, particles in at least a portion of the second test sample in the second reaction cell are passed one by one through an optically irradiated detection area to obtain the second optical information.

[0128] Further embodiments and advantages of the blood cell analysis method 200 proposed in this application can be found in the above description of the blood cell analyzer 100, and will not be repeated here.

[0129] All features or combinations of features mentioned above in the specification, drawings, and claims may be used in any combination or individually, provided they are meaningful within the scope of this application and do not contradict each other. The advantages and features described in the blood cell analyzer provided with reference to the embodiments of this application are applicable in a corresponding manner to the blood cell analysis method provided with the embodiments of this application, and vice versa.

[0130] The above description is merely a preferred embodiment of this application and does not limit the patent scope of this application. All equivalent modifications made based on the inventive concept of this application and the contents of the specification and drawings of this application, or direct / indirect applications in other related technical fields, are included within the patent protection scope of this application.

Claims

1. A blood cell analyzer, comprising: A sampling device is used to collect blood samples for testing. A sample preparation apparatus is used to mix a portion of the blood sample to be tested, a diluent, and a fluorescent staining agent in a first reaction cell to prepare a first test sample, and to mix another portion of the blood sample to be tested and a hemolysin in a second reaction cell to prepare a second test sample; An optical detection device includes a liquid driving unit, a flow chamber, a light source, and a photodetector. The liquid driving unit is used to drive the first test sample from the first reaction cell through the flow chamber. The light source is used to illuminate the first test sample passing through the flow chamber. The photodetector is used to detect the first optical information generated by the first test sample after being illuminated by light when passing through the flow chamber. A colorimetric detection device is used to detect the second test sample in the second reaction cell to obtain colorimetric optical information; The data processing device is configured as follows: Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained. The first white blood cell detection results include at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes. The granulocytes include neutrophils and eosinophils. as well as The hemoglobin concentration of the blood sample to be tested is obtained based on the colorimetric optical information.

2. The blood cell analyzer according to claim 1, characterized in that, The sample preparation device is further configured to add a chemical staining agent when preparing the first test sample, so that the data processing device can distinguish eosinophils in the granulocytes based on the first optical information, so that the first white blood cell detection result includes at least the percentage of lymphocytes, the percentage of monocytes, the percentage of eosinophils and the percentage of neutrophils. The data processing device is further configured to identify basophils in the first test sample based on the first optical information, such that the first white blood cell detection result also includes the percentage of basophils.

3. The blood cell analyzer according to claim 2, characterized in that, The sample preparation apparatus is further configured to add the chemical staining agent, either mixed in with the diluent and / or the fluorescent staining agent, or add it separately when preparing the first test sample.

4. The blood cell analyzer according to claim 2 or 3, characterized in that, The chemical dye is selected from acidic organic pigments; The chemical dye is selected from the Acid Blue series; The chemical dyes are selected from the group consisting of Direct Blue, Acid Green, Acid Yellow, Acid Orange, Methyl Red, Methyl Orange, Aniline Blue, Alizarin Yellow, Reactive Black, Sudan Black B, and Azo Black E.

5. The blood cell analyzer according to any one of claims 1 to 4, characterized in that, The data processing device acquires the reticulocyte detection results, platelet detection results, red blood cell detection results, and white blood cell detection results of the blood sample to be tested based on the first optical information, including: the data processing device A first scatter plot is generated based on at least the forward-scattered light signal and fluorescence signal in the first optical information, and the reticulocyte detection results, platelet detection results, and red blood cell detection results are obtained based on the first scatter plot; and A second scatter plot is generated based on at least the forward-scattered light signal and the side-scattered light signal in the first optical information, and the first white blood cell detection result is obtained based on the second scatter plot.

6. The blood cell analyzer according to any one of claims 1 to 5, characterized in that, The liquid driving unit is further configured to drive at least a portion of the second test sample from the second reaction cell through the flow chamber, the light source is further configured to irradiate the at least a portion of the second test sample passing through the flow chamber with light, and the photodetector is further configured to detect the second optical information generated after the at least a portion of the second test sample is irradiated with light when passing through the flow chamber. The data processing device is further configured to acquire a second white blood cell detection result of the blood sample to be tested based on the second optical information, the second white blood cell detection result including at least white blood cell count, basophil count and basophil percentage.

7. The blood cell analyzer according to claim 6, characterized in that, The data processing device acquires the second white blood cell detection result of the blood sample to be tested based on the second optical information, including: the data processing device A third scatter plot is generated based on the forward-scattered light signal and the side-scattered light signal in the second optical information, and the second white blood cell detection result is obtained based on the third scatter plot.

8. The blood cell analyzer according to claim 6 or 7, characterized in that, The liquid driving unit is configured to drive at least a portion of the second measurement sample from the second reaction cell through the flow chamber before or after the colorimetric detection device detects the second measurement sample in the second reaction cell to obtain the colorimetric optical information, so that the photodetector detects the second optical information.

9. A method for blood cell analysis, comprising: Collect the blood sample to be tested; A portion of the blood sample to be tested, a diluent, and a fluorescent staining agent are mixed in a first reaction cell to prepare a first test sample. Particles in the first test sample in the first reaction cell are then passed one by one through an optically irradiated detection area to obtain the first optical information generated by the particles in the first test sample after being irradiated by light. Another portion of the blood sample to be tested and a hemolysin are mixed in a second reaction chamber to prepare a second test sample, and the second test sample in the second reaction chamber is detected by colorimetry to obtain colorimetric optical information. Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained. The first white blood cell detection results include at least the percentage of lymphocytes, the percentage of monocytes, and the percentage of granulocytes. The granulocytes include neutrophils and eosinophils. as well as The hemoglobin concentration of the blood sample to be tested is determined based on the colorimetric optical information.

10. The blood cell analysis method according to claim 9, characterized in that, In the preparation of the first test sample, a chemical staining agent is also added to distinguish eosinophils in the granulocytes based on the first optical information, so that the first white blood cell test result includes at least the percentage of lymphocytes, the percentage of monocytes, the percentage of eosinophils and the percentage of neutrophils. Based on the first optical information, basophils in the first test sample are also identified, so that the first white blood cell detection result also includes the percentage of basophils.

11. The blood cell analysis method according to claim 10, characterized in that, The chemical staining agent is added either by mixing it into the diluent and / or the fluorescent staining agent or by adding it alone when preparing the first test sample.

12. The blood cell analysis method according to claim 10 or 11, characterized in that, The chemical dye is selected from acidic organic pigments; The chemical dye is selected from the Acid Blue series; The chemical dyes are selected from the group consisting of Direct Blue, Acid Green, Acid Yellow, Acid Orange, Methyl Red, Methyl Orange, Aniline Blue, Alizarin Yellow, Reactive Black, Sudan Black B, and Azo Black E.

13. The blood cell analysis method according to any one of claims 9 to 12, characterized in that, Based on the first optical information, the reticulocyte detection results, platelet detection results, red blood cell detection results, and first white blood cell detection results of the blood sample to be tested are obtained, including: A first scatter plot is generated based on at least the forward-scattered light signal and fluorescence signal in the first optical information, and the reticulocyte detection results, platelet detection results, and red blood cell detection results are obtained based on the first scatter plot; and A second scatter plot is generated based on at least the forward-scattered light signal and the side-scattered light signal in the first optical information, and the first white blood cell detection result is obtained based on the second scatter plot.

14. The blood cell analysis method according to any one of claims 9 to 13, further comprising: The particles in at least a portion of the second test sample in the second reaction cell are passed one by one through an optical detection area irradiated by light to obtain the second optical information generated by the particles in at least a portion of the second test sample after being irradiated by light. as well as The second white blood cell detection result of the blood sample to be tested is obtained based on the second optical information. The second white blood cell detection result includes at least the white blood cell count, basophil count and basophil percentage.

15. The blood cell analysis method according to claim 14, characterized in that, The second white blood cell detection result of the blood sample to be tested is obtained based on the second optical information, including: A third scatter plot is generated based on the forward-scattered light signal and the side-scattered light signal in the second optical information, and the second white blood cell detection result is obtained based on the third scatter plot.

16. The blood cell analysis method according to claim 14 or 15, characterized in that, Before or after the second test sample in the second reaction cell is tested by colorimetry to obtain the colorimetric optical information, particles in at least a portion of the second test sample in the second reaction cell are passed one by one through an optically irradiated detection area to obtain the second optical information.

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