Sample analyzer and sample analysis method

Abstract

The embodiment of the invention relates to a sample analyzer and a sample analysis method. The sample analysis method comprises the following steps: sucking a blood sample to be detected; mixing at least one part of the blood sample to be detected, the hemolytic agent, the first fluorescent dye and the second fluorescent dye to prepare a detection sample; in a single test, each particle in a test sample passes through an optical detection area irradiated by exciting light to obtain optical information generated after each particle in the test sample is irradiated by the exciting light, and the exciting light comprises 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 greater than the first wavelength; obtaining infected red blood cell information of the test sample based on the at least one scattered light signal and the first fluorescence signal; and based on the at least one scattered light signal and the second fluorescence signal, obtaining a leukocyte four-classification result of the determination sample. Therefore, the infected red blood cell information and the white blood cell classification result of the test sample can be simultaneously obtained in one test.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of blood cell detection, and in particular to a sample analyzer and a sample analysis method. BACKGROUND

[0002] Malaria is a serious parasitic disease that seriously endangers human health caused by Plasmodium parasitizing in human body. It is mainly transmitted in humans and mosquitoes by female Anopheles biting humans infected with Plasmodium, and is one of the most serious tropical insect-borne infectious diseases in the world so far, with a high mortality rate. If malaria is not timely differentiated from other febrile diseases for treatment, malaria can develop into a serious disease within 24 hours, causing illness delay, leading to severe and death cases.

[0003] In the related art, blood cell counting using a blood cell analyzer is a routine detection item for patients with fever.

[0004] With the development of blood cell analysis technology in recent years, blood cell analyzers have developed from single-cell counting detection to multi-functional joint detection instruments that can simultaneously analyze tens of thousands of cells at high speed and obtain multiple parameters describing cell morphological characteristics. Using digital and image results in blood analyzers helps to screen for malaria, but the application of blood cell analyzers in the related art in malaria diagnosis still has many limitations. SUMMARY

[0005] One task of the present application is to obtain infected red blood cell information and white blood cell classification results through only one reaction and detection channel.

[0006] Another task of the present application is to obtain infected red blood cell information and nucleated red blood cell information through only one reaction and detection channel.

[0007] To achieve the tasks of the present application, the first aspect of the present application relates to a sample analyzer, comprising:

[0008] a sample suction device for sucking a blood sample to be tested;

[0009] a sample preparation device for mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a second fluorescent dye in a reaction pool to prepare a test sample;

[0010] An optical detection device comprises a light emitting device for emitting excitation light to irradiate a flow cell, a flow chamber in communication with the flow cell and for passing each particle in the assay sample therethrough, and a light detector for detecting optical information generated by each particle in the assay sample when irradiated by the excitation light in a single test, the light emitting device comprising a first light source for emitting light of a first wavelength capable of exciting the first fluorescent dye and a second light source for emitting light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a second fluorescent signal corresponding to the second fluorescent dye detected in the single test; and

[0011] a processor configured to obtain infected red blood cell information of the assay sample based on at least one of the scatter light signals and the first fluorescent signal, and obtain a white blood cell four-classification result of the assay sample based on at least one of the scatter light signals and the second fluorescent signal, wherein the white blood cell four-classification result comprises a count result of a lymphocyte population, a count result of a monocyte population, a count result of a neutrophil population, and a count result of an eosinophil population in the assay sample.

[0012] To achieve the object of the present application, the second aspect of the present application relates to a sample analysis method, comprising:

[0013] aspirating a blood sample to be tested;

[0014] mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye to prepare an assay sample;

[0015] passing each particle in the assay sample through an optical detection region irradiated by excitation light in a single test to obtain optical information generated by each particle in the assay sample when irradiated by the excitation light, the excitation light comprising light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a second fluorescent signal corresponding to the second fluorescent dye obtained in the single test;

[0016] obtaining infected red blood cell information of the assay sample based on at least one of the scatter light signals and the first fluorescent signal, and

[0017] The white blood cell four-part differential result of the test sample is obtained based on at least one of the scattered light signals and the second fluorescence signal, wherein the white blood cell four-part differential result includes the count results of lymphocytes, monocytes, neutrophils and eosinophils in the test sample.

[0018] In the technical solutions provided in the first and second aspects of this application, a blood sample to be tested, a hemolysin, and two fluorescent dyes are mixed in a reaction and detection channel to prepare a test sample, and the test sample is optically measured in one test to obtain information on infected red blood cells and white blood cell four-part differential results based on the optical information obtained in one test.

[0019] To achieve the objectives of this application, a third aspect of this application relates to a sample analyzer, comprising:

[0020] A sampling device is used to collect blood samples for testing.

[0021] A sample preparation apparatus for mixing at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample;

[0022] An optical detection device includes a light emitting device, a flow chamber, and a photodetector. The light emitting device emits excitation light to irradiate the flow chamber, which is connected to a reaction cell and allows particles in the test sample to pass through. The photodetector detects the optical information generated when particles in the test sample are irradiated by the excitation light as they pass through the flow chamber in a single test. The light emitting device includes a first light source and a second light source. The first light source emits light of a first wavelength capable of exciting a first fluorescent dye, and the second light source emits light of a second wavelength capable of exciting a third fluorescent dye. Either the first or second wavelength light can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in the single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0023] The processor is configured to obtain information on infected red blood cells and white blood cell differential counts of the test sample based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal, and the third fluorescence signal, wherein the white blood cell differential counts include the counts of lymphocytes, monocytes, neutrophils, eosinophils, and basophils in the test sample.

[0024] To achieve the objectives of this application, a fourth aspect of this application relates to a sample analysis method, comprising:

[0025] Collect the blood sample to be tested;

[0026] The test sample is prepared by mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye.

[0027] In a single test, each particle in the test sample is passed through an optical detection area irradiated by excitation light to obtain optical information generated by each particle after being irradiated by the excitation light. The excitation light includes light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the third fluorescent dye. Either the first wavelength or the second wavelength can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal obtained in the single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye; and

[0028] The infected red blood cell information and white blood cell five-part differential results of the test sample are obtained based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, wherein the white blood cell five-part differential results include the count results of lymphocyte population, monocyte population, neutrophil population, eosinophil population and basophil population in the test sample.

[0029] In the technical solutions provided in the third and fourth aspects of this application, a blood sample to be tested, a hemolysin, and three fluorescent dyes are mixed in a reaction and detection channel to prepare a test sample, and the test sample is optically measured in one test to obtain information on infected red blood cells and white blood cell five-part differential results based on the optical information obtained in one test.

[0030] To achieve the objectives of this application, a fifth aspect of this application relates to a sample analyzer, comprising:

[0031] A sampling device is used to collect blood samples for testing.

[0032] A sample preparation apparatus for mixing at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample;

[0033] An optical detection device includes a light emitting device, a flow chamber, and a photodetector. The light emitting device emits excitation light to irradiate the flow chamber, which is connected to a reaction cell and allows individual particles in the test sample to pass through. The photodetector detects, in a single test, the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow chamber. The optical information includes at least one scattered light signal detected in the single test, a first fluorescence signal corresponding to a first fluorescent dye, and a third fluorescence signal corresponding to a third fluorescent dye; and

[0034] The processor is configured to obtain information on infected red blood cells and nucleated red blood cells of the test sample based on at least one of the scattered light signals, the first fluorescence signal, and the third fluorescence signal.

[0035] To achieve the objectives of this application, the sixth aspect of this application relates to a sample analysis method, comprising:

[0036] Collect the blood sample to be tested;

[0037] The test sample is prepared by mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye.

[0038] In a single test, each particle in the test sample is passed through an optical detection area irradiated by excitation light to obtain optical information generated by each particle after being irradiated by the excitation light. This optical information includes at least one scattered light signal obtained in the single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye; and

[0039] The information on infected red blood cells and nucleated red blood cells of the test sample is obtained based on at least one of the scattered light signals, the first fluorescence signal, and the third fluorescence signal.

[0040] In the technical solutions provided in the fifth and sixth aspects of this application, a blood sample to be tested, a hemolysin, and two dyes are mixed in a reaction and detection channel to prepare a test sample, and the test sample is optically measured in one test to obtain information on infected red blood cells and nucleated red blood cells based on the optical information obtained in one test. Attached Figure Description

[0041] Figure 1 This is a schematic diagram of one embodiment of the sample analyzer according to this application.

[0042] Figure 2 This is a schematic diagram of one embodiment of the optical inspection device according to this application.

[0043] Figure 3 This is a schematic flowchart of one embodiment of the sample analysis method according to this application.

[0044] Figure 4 This is a schematic flowchart of another embodiment of the sample analysis method according to this application.

[0045] Figure 5 This is a schematic flowchart of yet another embodiment of the sample analysis method according to this application.

[0046] Figure 6a , Figure 6b and Figure 6c This is a scatter plot obtained from testing a blood sample that is not infected with malaria parasites according to the first embodiment of this application.

[0047] Figure 7a , Figure 7b and Figure 7c This is a scatter plot obtained from the detection of a blood sample infected with malaria parasites according to the first embodiment of this application.

[0048] Figure 8a , Figure 8b and Figure 8c This is a scatter plot obtained from the detection of the blood sample to be tested according to the second embodiment of this application.

[0049] Figure 9a , Figure 9b and Figure 9c This is a scatter plot obtained from the detection of the blood sample to be tested according to the third embodiment of this application.

[0050] Figure 10a , Figure 10b , Figure 10c and Figure 10d This is a scatter plot obtained from the detection of the blood sample to be tested according to the fourth embodiment of this application.

[0051] Figure 11a and Figure 11b This is a scatter plot obtained from the detection of the blood sample to be tested according to the fifth embodiment of this application. Detailed Implementation

[0052] The embodiments of this application will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are merely a part of the embodiments of this application, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

[0053] The serial numbers assigned to components in this document, such as "first" and "second," are used only to distinguish the described objects and have no sequential or technical meaning. The terms "connection" and "linkage" used in this application, unless otherwise specified, include both direct and indirect connections (linkages).

[0054] To facilitate subsequent explanations, a brief explanation of some terms used in this application is provided below:

[0055] 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.

[0056] The term "scatter plot" as used in this application refers not only to a distribution of at least two sets of data in a rectangular coordinate system in the form of data points, but also to data arrays, i.e., regardless of the form in which they are presented graphically.

[0057] 2) Cell clusters: A group of particles with the same characteristics distributed in a certain area of ​​a scatter plot. It is also called a "particle cluster" or "cell cluster". For example, white blood cell (including all types of white blood cells) clusters, as well as white blood cell subpopulations, such as neutrophil clusters, lymphocyte clusters, monocyte clusters, eosinophil clusters, or basophil clusters.

[0058] 3) Blood shadow: Fragment particles obtained by dissolving red blood cells and platelets in blood with a hemolytic agent.

[0059] The blood cell analyzer used in this application classifies and counts particles in blood samples using flow cytometry combining laser scattering and fluorescence staining. The principle of the blood cell analyzer in detecting blood samples can be illustrated as follows: First, a blood sample is drawn and treated with a hemolysin and fluorescent dye. Red blood cells are destroyed and dissolved by the hemolysin, while white blood cells are not dissolved. However, the fluorescent dye can enter the nucleus of white blood cells with the help of the hemolysin and bind to nucleic acid substances in the nucleus. Next, each particle in the sample passes through a detection aperture illuminated by a laser beam. When the laser beam illuminates the particles, the characteristics of the particles themselves (such as volume, staining degree, size and content of cell contents, and nuclear density) can block or change the direction of the laser beam, thereby generating scattered light at various angles corresponding to the particle characteristics. This scattered light is received by a signal detector to obtain 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 light information can be used to classify and count cells in blood samples.

[0060] Figure 1 This is a schematic diagram of the structure of a sample analyzer according to some embodiments of this application. The sample analyzer 100 includes at least a sample aspiration device 110, a sample preparation device 120, an optical detection device 130, and a processor 140. The sample aspiration device 110 is used to aspirate a blood sample to be tested. The sample preparation device 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, and at least two fluorescent dyes in a reaction cell to prepare a test sample. The optical detection device 130 includes a light emitting device, a flow cell, and a photodetector. The light emitting device emits excitation light to irradiate the flow cell, which is connected to the reaction cell and allows individual particles in the test sample to pass through. The photodetector detects, in a single test, the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow cell. The optical information includes at least one scattered light signal and at least two fluorescent signals detected in a single test.

[0061] 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 to be tested from the test tube through the tip of the sampling needle. Additionally, the sampling device 110 can also deliver the drawn-up blood sample to the sample preparation device 120.

[0062] In some embodiments, the sample preparation apparatus 120 may include at least one reaction cell and a reagent supply device (not shown). The at least one 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 dye, etc.) to the at least one 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 reaction cell to prepare a test sample.

[0063] For example, the sample preparation device 120 is used to mix and incubate blood samples, hemolysin, and at least two fluorescent dyes for a reaction time of 10 s to 1 min, a reaction temperature of 25 ℃ to 50 ℃, and a reaction time of 10 s to 40 s.

[0064] In some embodiments, the sample preparation apparatus 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a second fluorescent dye in a reaction cell to prepare a test sample.

[0065] In other embodiments, the sample preparation apparatus 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample.

[0066] In some other embodiments, the sample preparation apparatus 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample.

[0067] In this embodiment, the hemolytic agent can dissolve red blood cells and platelets in a blood sample, but can maintain the morphology of white blood cells essentially unchanged. The fragment particles obtained after the red blood cells and platelets in the blood sample are dissolved form a blood shadow, thereby distinguishing it from the white blood cells and nucleated red blood cells in the blood sample.

[0068] In some embodiments, the hemolytic agent may comprise one or more combinations of cationic surfactants, nonionic surfactants, anionic surfactants, amphiphilic surfactants, and buffer pairs. Cationic surfactants are, for example, selected from at least one or more combinations of dodecyltrimethylammonium chloride, octyltrimethylammonium bromide, and tetradecyltrimethylammonium chloride. Nonionic surfactants are, for example, selected from at least one or more combinations of long-chain fatty alcohol polyoxyethylene, alkylphenol polyoxyethylene ethers, fatty acid polyoxyethylene ethers, and fatty amine polyoxyethylene ethers. Buffer pairs are, for example, selected from at least one or more combinations of phosphates, citrates, Tris-HCl, and MES buffers. Anionic surfactants are, for example, selected from at least one or more combinations of dodecylbenzenesulfonic acid, sodium fatty alcohol acyl sulfate, ethoxylated fatty acid methyl ester sulfonate sodium, secondary alkyl sulfonate sodium, and alcohol ether carboxylates.

[0069] In some embodiments, the hemolytic agent may include a buffer solution that can adjust the pH of the hemolytic agent to between 5 and 11, preferably between 6 and 8.

[0070] In other embodiments, the hemolytic agent may include at least one of alkyl glycosides, triterpenoid saponins, and steroidal saponins.

[0071] In this embodiment, the hemolytic agent can enhance the permeability of blood cells with a short hemolysis time, allowing the fluorescent dye to enter the cells and specifically bind and stain them. Because the binding abilities of nucleic acids in leukocytes and nucleated erythrocytes, as well as cytochromes and riboflavin within erythrocytes and platelets, to the fluorescent dye differ, different fluorescent signals are generated.

[0072] In some embodiments, the first fluorescent dye may be a fluorescent dye capable of specifically labeling malaria parasites, comprising compounds with the following general structural formula:

[0073]

[0074] In this general formula,

[0075] R1 is selected from substituted or unsubstituted phenyl groups;

[0076] R2 is selected from C 1-8 Nitrogen-containing alkyl groups, substituted or unsubstituted nitrogen- or oxygen-containing heterocycles; wherein the nitrogen- or oxygen-containing heterocycle is substituted or unsubstituted phenyl groups;

[0077] X is a halogen or PF6. - ;

[0078] The phenyl groups of R1 and R2 are each independently selected from the following substituents: CN, COOH, NH2, NO2, OH, SH, C. 1-6 Alkoxy, C 1-6 Alkylamino, C 1-6 Amide group, halogen or C 1-6 Halogenated alkyl groups.

[0079] Furthermore, R1 is selected from hydrogen, phenyl, or biphenyl. Even further, R1 is selected from biphenyl.

[0080] Further, R2 is selected from N,N-dimethylpiperazinyl or 2,6-dimethylpiperazinyl. Even further, R2 is selected from 2,6-dimethylpiperazinyl.

[0081] As some embodiments, the compound used for the first fluorescent dye has one of the following structures: A, B, C, and D:

[0082] Compound A

[0083]

[0084] Compound B

[0085]

[0086] Compound C

[0087]

[0088] Compound D

[0089]

[0090] Here, the concentration of the first fluorescent dye is 0.1-1000 mg / L.

[0091] Further embodiments of the first fluorescent dye of this application can be found in Chinese application CN202310028120.1 filed by the applicant on January 9, 2023, the entire disclosure of which is incorporated herein by reference.

[0092] In some embodiments, the second fluorescent dye may be a fluorescent dye capable of being used for leukocyte classification, comprising compounds with the following general structural formula:

[0093]

[0094] Here, the concentration of the second fluorescent dye is 0.1-1000 mg / L.

[0095] Further embodiments of the second fluorescent dye of this application can be found in the applicant's U.S. Patent US7598385B2, the entire disclosure of which is incorporated herein by reference.

[0096] In some embodiments, the third fluorescent dye may be a fluorescent dye capable of recognizing nucleated erythrocytes, comprising compounds with the following general structural formula:

[0097] In this general formula, X is selected from -O-, -S-, and One of them; R1 and R2 are each independently selected from C3-C 20 alkenyl and C3-C 20 Alkynes.

[0098] As some implementations, compounds used for third fluorescent dyes have one of the following structures: I, II, III, and IV:

[0099] Compound I

[0100]

[0101] Compound II

[0102]

[0103] Compound III

[0104]

[0105] Compound IV

[0106]

[0107] Compound V

[0108]

[0109] Here, the concentration of the third fluorescent dye is 0.1-1000 mg / L.

[0110] In some embodiments, the first fluorescent dye and / or the second fluorescent dye and / or the third fluorescent dye are stored in a water-soluble organic phase such as glycerol, glycol, or ethylene glycol.

[0111] In some embodiments, the first fluorescent dye and / or the second fluorescent dye and / or the third fluorescent dye may be stored separately or mixed with a hemolysin.

[0112] In some embodiments, the light emitting device includes a first light source and a second light source. The first light source is used to emit light of a first wavelength capable of exciting a first fluorescent dye, and the second light source is used to emit light of a second wavelength capable of exciting a second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a second fluorescence signal corresponding to the second fluorescent dye.

[0113] In other embodiments, the light emitting device includes a first light source and a second light source. The first light source emits light of a first wavelength capable of exciting a first fluorescent dye, and the second light source emits light of a second wavelength capable of exciting a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0114] In yet other embodiments, the light emitting device includes a first light source and a second light source. The first light source emits light of a first wavelength capable of exciting a first fluorescent dye and a second dye, and the second light source emits light of a second wavelength capable of exciting a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0115] In some further embodiments, the light emitting device includes a first light source and a second light source. The first light source emits light of a first wavelength capable of exciting a first fluorescent dye, and the second light source emits light of a second wavelength capable of exciting a second and a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0116] In some embodiments, the flow chamber in the optical detection device 130 refers to a chamber with a focused liquid flow suitable for detecting scattered light signals and fluorescence signals. When a particle, such as a blood cell, passes through the detection aperture of the flow chamber, the particle scatters the incident light beam from the light source, which is guided through the detection 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 scattered light pulse signal (also known as a scattered light signal). Since different particles have different light scattering characteristics, the scattered light signal can be used to distinguish different groups of particles.

[0117] Specifically, the scattered light signal detected near the incident light beam is generally referred to as the forward scattered light signal or the small-angle scattered light signal. In some embodiments, the forward scattered light 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 scattered light signal can be detected at an angle of about 2° to about 6° with respect to the incident light beam. The scattered light signal detected at about 90° with respect to the incident light beam is generally referred to as the side scattered light signal. In some embodiments, the side scattered light 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.

[0118] In some embodiments, the photodetector in the optical detection device 130 may include a scattering light detector for detecting scattered light pulse signals (also known as scattered light signals), such as side-scattered light pulse signals (also known as side-scattered light signals), and a first fluorescence detector for detecting a first fluorescence pulse signal (also known as a first fluorescence signal) and a second fluorescence detector for detecting a second fluorescence pulse signal (also known as a second fluorescence signal).

[0119] In other embodiments, the photodetector in the optical detection device 130 may include a scattering light detector for detecting scattered light pulse signals (also known as scattered light signals), such as side-scattered light pulse signals (also known as side-scattered light signals), and a first fluorescence detector for detecting a first fluorescence pulse signal (also known as a first fluorescence signal) and a third fluorescence detector for detecting a third fluorescence pulse signal (also known as a third fluorescence signal).

[0120] In some other embodiments, the photodetector in the optical detection device 130 may include a scattering light detector for detecting scattered light pulse signals (also known as scattered light signals), such as side-scattered light pulse signals (also known as side-scattered light signals), and a first fluorescence detector for detecting a first fluorescence pulse signal (also known as a first fluorescence signal), a second fluorescence detector for detecting a second fluorescence pulse signal (also known as a second fluorescence signal), and a third fluorescence detector for detecting a third fluorescence pulse signal (also known as a third fluorescence signal).

[0121] In some embodiments, the optical detection device 130 includes a forward-scattering detector for detecting forward-scattered light signals or a side-scattering detector for detecting side-scattered light signals. Preferably, the optical detection device 130 includes both a forward-scattering detector and a side-scattering detector.

[0122] Figure 2This is a specific example of the optical detection device 130 of this application. The optical detection device 130 includes a light emitting device 131, a front light assembly 132, a flow chamber 133, a forward scattering light detector 134, a first dichroic mirror 135, a side scattering light detector 136, a second dichroic mirror 137, a first fluorescence detector 138, and a second fluorescence detector 139. The first fluorescence detector 138 is used to detect a first fluorescence signal corresponding to a first fluorescent dye generated by particles passing through the flow chamber 133 after being irradiated by a light beam. The second fluorescence detector 139 is used to detect a second fluorescence signal corresponding to a second fluorescent dye generated by particles passing through the flow chamber 133 after being irradiated by a light beam. Here, the light emitting device 131, the front light assembly 132, the flow chamber 133, and the forward scattering light detector 134 are arranged sequentially along the optical axis. The front light assembly 132 is configured to converge the light beam emitted by the light emitting device 131 into the detection area of ​​the flow chamber 133 in the particle flow direction, so that particles flowing through the detection area of ​​the flow chamber 133 can generate scattered light. On one side of the flow chamber 133, a first dichroic mirror 135 is arranged at a 45° angle to the optical axis. A portion of the lateral light generated when particles flow through the detection area of ​​the flow chamber 133 is reflected by the first dichroic mirror 135 and captured by the side-scattering light detector 136, while another portion of the lateral light passes through the first dichroic mirror 135 and reaches a second dichroic mirror 137, which is also arranged downstream of the first dichroic mirror 135 at a 45° angle to the optical axis. A portion of the lateral light passing through the first dichroic mirror 135 is reflected by the second dichroic mirror 137 and captured by the first fluorescence detector 138, while another portion passes through the second dichroic mirror 137 and is captured by the second fluorescence detector 139.

[0123] In some embodiments, the first wavelength is between 315 nm and 490 nm and the second wavelength is between 610 nm and 750 nm. Preferably, the first wavelength is between 400 nm and 450 nm and the second wavelength is between 620 nm and 700 nm. That is, the first wavelength of light includes blue-violet light and the second wavelength of light includes red light.

[0124] In some embodiments, the processor 140 is used to process and perform calculations on the data to obtain the desired results, such as generating a two-dimensional or three-dimensional scatter plot based on the collected optical information, and performing particle analysis on the scatter plot according to the gating method.

[0125] In some embodiments, the processor 140 may visualize intermediate or final processing results and then display them through the display device 150. For example, the display device 150 may include a user interface, and the processor 140 may display the processing results on the user interface of the display device 150.

[0126] In some embodiments, the processor 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 processor (DSP) for interpreting computer instructions and processing data in computer software. For example, the processor is used to execute various computer applications in a computer-readable storage medium, thereby enabling the sample analyzer 100 to execute corresponding detection procedures and analyze the scattered light signal and fluorescence signal detected by the optical detection device 130 in real time.

[0127] In addition, the sample analyzer 100 may also include a liquid path system (not shown) for connecting the sample aspiration device 110, the sample preparation device 120 and the optical detection device 130 to facilitate liquid transfer between these devices.

[0128] Furthermore, the sample analyzer 100 may also include a first housing 160 and a second housing 170. An optical detection device 130 and a processor 140 are disposed inside the second housing 170. A sample preparation device 120 is disposed, for example, inside the first housing 160, and a 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 sample analyzer 100.

[0129] In a first embodiment of this application, the sampling device 110 is used to aspirate a blood sample to be tested; the sample preparation device 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a second fluorescent dye in a reaction cell to prepare a test sample; the optical detection device 130 includes a light emitting device, a flow cell, and a photodetector. The light emitting device is used to emit excitation light to irradiate the flow cell. The flow cell is connected to the reaction cell and is used to allow individual particles in the test sample to pass through. The photodetector is used to detect the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow cell in a single test. The light emitting device includes a first light source and a second light source. The first light source is used to emit light of a first wavelength that can excite the first fluorescent dye, and the second light source is used to emit light of a second wavelength that can excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a second fluorescence signal corresponding to the second fluorescent dye. Here, the processor 140 is configured to: obtain infected erythrocyte information of the test sample based on at least one scattered light signal and a first fluorescence signal detected in a single test of a test sample prepared by mixing a blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye; and obtain a four-part differential white blood cell count of the test sample based on at least one scattered light signal and a second fluorescence signal detected in the single test, wherein the four-part differential white blood cell count includes the counts of lymphocytes (lym), monocytes (mon), neutrophils (neu), and eosinophils (eos) in the test sample. Here, infected erythrocyte information refers to optical information related to infected erythrocytes.

[0130] This enables the acquisition of white blood cell four-part differential results and information on infected red blood cells through a single reaction and detection channel. In other words, it allows for the specific detection of red blood cells infected with malaria parasites while simultaneously classifying white blood cells, saving detection time and blood volume, and achieving efficient diagnosis of the disease status of subjects (e.g., patients with fever).

[0131] In some embodiments, the scattered light signal includes: a first forward scattered light signal and a first side scattered light signal generated by particles passing through the flow chamber and irradiated with light of a first wavelength in a single test, and / or a second forward scattered light signal and a second side scattered light signal generated by particles passing through the flow chamber and irradiated with light of a second wavelength in a single test. The processor 140 obtains information on infected red blood cells in the test sample based on at least one scattered light signal and a first fluorescence signal, including: generating a first scatter plot based on one of the first forward scattered light signal, the first side scattered light signal, the second forward scattered light signal, the second side scattered light signal, and the first fluorescence signal, and obtaining information on infected red blood cells in the test sample based on the first scatter plot.

[0132] In some embodiments, infected red blood cell information may include a count of infected red blood cells. Obtaining quantitative results for infected red blood cell detection can more accurately assist doctors in diagnosing malaria.

[0133] In other embodiments, the information about infected red blood cells may include the type of Plasmodium parasite infecting the red blood cells. Typing the Plasmodium parasites infecting the red blood cells can assist doctors in diagnosing malaria and enabling effective treatment.

[0134] In some embodiments, the processor 140 obtains information on infected red blood cells in the test sample based on a first scatter plot, including:

[0135] The count of infected red blood cells in the test sample was obtained based on the first scatter plot; and / or

[0136] The type of malaria parasite infecting the infected red blood cells in the test sample is determined based on the first scatter plot.

[0137] In a specific example, a first scatter plot is generated based on a first forward-scattered light signal and a first fluorescence signal. Using a gating technique, a region representing infected red blood cells is obtained from this first scatter plot. The scatter points falling into this region are counted to obtain the count result of infected red blood cells, i.e., the count value. If the count value of infected red blood cells is greater than a predetermined threshold, an alarm is output (indicating that the blood sample is a malaria-positive sample).

[0138] In another specific example, a first scatter plot is generated based on a first forward-scattered light signal and a first fluorescence signal. Using a gating technique, regions representing infected red blood cells are obtained from this first scatter plot. From these regions, a first infected red blood cell population and a second infected red blood cell population are identified. Each infected red blood cell in the second infected red blood cell population contains multiple Plasmodium rings. The centroids of the first fluorescence signal and the first forward-scattered light signal of the second infected red blood cell population are correspondingly larger than those of the first fluorescence signal and the first forward-scattered light signal of the first infected red blood cell population. This allows for a more accurate acquisition of the infected red blood cell count, avoiding the misidentification of a single Plasmodium ring within multiple Plasmodium rings in an infected red blood cell as a single infected red blood cell.

[0139] In some embodiments, the processor 140 generates a first scatter plot based on one of the following signals: a first forward-scattered light signal, a first side-scattered light signal, a second forward-scattered light signal, a second side-scattered light signal, and a first fluorescence signal. This includes generating the first scatter plot based on either the first forward-scattered light signal and the first fluorescence signal, or based on either the second forward-scattered light signal and the first fluorescence signal. Here, the processor 140 determines the type of Plasmodium infected by the infected red blood cells in the test sample based on the first scatter plot. This includes identifying infected red blood cell regions containing infected red blood cell clusters from the first scatter plot, and determining, based on the forward-scattered light intensity of the infected red blood cell clusters in the infected red blood cell regions, whether the Plasmodium infected by the infected red blood cells in the test sample is Plasmodium falciparum, Plasmodium ovale, or Plasmodium vivax.

[0140] In a specific example, a first scatter plot is generated based on a first forward-scattered light signal and a first fluorescence signal. Using a gating technique, a region representing infected erythrocytes is obtained from this first scatter plot. The type of Plasmodium infected by the erythrocytes is determined by the magnitude of the centroid or peak value of the first forward-scattered light signal within this region. Because different types of Plasmodium differ in size, the intensity of the first forward-scattered light signal differs between erythrocytes infected with *Plasmodium falciparum*, *Plasmodium ovale*, and *Plasmodium vivax*. Specifically, the centroid of the first forward-scattered light signal of erythrocytes infected with *Plasmodium vivax* is larger than that of erythrocytes infected with *Plasmodium ovale*, and the centroid of the first forward-scattered light signal of erythrocytes infected with *Plasmodium ovale* is larger than that of erythrocytes infected with *Plasmodium falciparum*.

[0141] In some embodiments, the scattered light signal includes: a first lateral scattered light signal generated by particles passing through the flow chamber and irradiated with light of a first wavelength, detected in a single test, and / or a second lateral scattered light signal generated by particles passing through the flow chamber and irradiated with light of a second wavelength, detected in a single test. Here, the processor 140 obtains the white blood cell four-part differential result of the test sample based on at least one scattered light signal and a second fluorescence signal, including: generating a second scatter plot based on the first lateral scattered light signal and the second fluorescence signal, or based on the second lateral scattered light signal and the second fluorescence signal, and obtaining the white blood cell four-part differential result of the test sample based on the second scatter plot.

[0142] In a second embodiment of this application, the sampling device 110 is used to aspirate the blood sample to be tested; the sample preparation device 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample. The optical detection device 130 includes a light emitting device, a flow cell, and a photodetector. The light emitting device is used to emit excitation light to irradiate the flow cell. The flow cell is connected to the reaction cell and is used to allow individual particles in the test sample to pass through. The photodetector is used to detect the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow cell in a single test. The light emitting device includes a first light source and a second light source. The first light source is used to emit light of a first wavelength capable of exciting the first fluorescent dye, and the second light source is used to emit light of a second wavelength capable of exciting the third fluorescent dye. The first wavelength light or the second wavelength light can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye. Here, the processor 140 is configured to obtain information on infected red blood cells and white blood cell differential counts of the test sample based on at least one scattered light signal, a first fluorescence signal, a second fluorescence signal, and a third fluorescence signal detected in a single test of a test sample prepared by mixing a blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye. The white blood cell differential counts include the counts of lymphocytes (lym), monocytes (mon), neutrophils (neu), eosinophils (eos), and basophils (baso) in the test sample.

[0143] In some embodiments, infected red blood cell information may include the count of infected red blood cells and / or the type of malaria parasite infecting the infected red blood cells.

[0144] In some embodiments, the processor 140 obtains information on infected red blood cells and white blood cell differential counts of the test sample based on at least one scattered light signal, a first fluorescence signal, a second fluorescence signal, and a third fluorescence signal, including: obtaining information on infected red blood cells based on one of at least one scattered light signal, a second fluorescence signal, a third fluorescence signal, and a first fluorescence signal; and obtaining white blood cell differential counts based on at least one scattered light signal, a second fluorescence signal, and a third fluorescence signal.

[0145] In some embodiments, the processor 140 obtains infected erythrocyte information based on at least one of a scattered light signal, a second fluorescence signal, a third fluorescence signal, and a first fluorescence signal, including: generating a first scatter plot based on at least one scattered light signal and the first fluorescence signal, and obtaining infected erythrocyte information based on the first scatter plot. The processor 140 obtains a five-part differential white blood cell count based on at least one scattered light signal, a second fluorescence signal, and a third fluorescence signal, including: generating a second scatter plot based on at least one scattered light signal and the second fluorescence signal, obtaining a four-part differential white blood cell count of the test sample based on the second scatter plot, wherein the four-part differential white blood cell count includes the count of lymphocytes, the count of monocytes, the count of neutrophils, and the count of eosinophils in the test sample; and identifying a first characteristic cell group including basophils in the second scatter plot, and obtaining the count of basophils in the test sample based on at least a third fluorescence signal of the first characteristic cell group.

[0146] In other embodiments, the processor 140 obtains a five-part differential white blood cell count based on at least one scattered light signal, a second fluorescence signal, and a third fluorescence signal, including: generating a second scatter plot based on at least one scattered light signal, such as a side-scattered light signal, and a second fluorescence signal, and obtaining a four-part differential white blood cell count of the test sample based on the second scatter plot; and generating a third scatter plot based on at least one scattered light signal, such as a side-scattered light signal, and a third fluorescence signal, and obtaining a count of basophils in the test sample based on the second scatter plot and the third scatter plot.

[0147] The inventors of this application discovered through research that nucleated erythrocytes and basophils can be accurately distinguished using scattered light signals, second fluorescence signals, and third fluorescence signals. Specifically, the processor 140 can distinguish between basophil populations and nucleated erythrocyte populations in the test sample based on at least one scattered light signal, second fluorescence signal, and third fluorescence signal, thereby identifying and counting the basophil populations in the test sample to obtain the count result of the basophil populations in the test sample.

[0148] In some implementations, the processor 140 obtains the basophil count in the test sample based on a second scatter plot and a third scatter plot, including: dividing the second scatter plot into a first characteristic cell group comprising basophils and nucleated erythrocytes and obtaining a first cell count of the first characteristic cell group; dividing the third scatter plot into a second characteristic cell group comprising only nucleated erythrocytes and obtaining a second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the basophil count of the test sample. That is, the first cell count is the count of basophils and nucleated erythrocytes in the test sample, and the second cell count is the count of nucleated erythrocytes in the test sample. Here, subtracting the second cell count from the first cell count yields the basophil count of the test sample.

[0149] Here, the second cell count is the count of nucleated red blood cells in the test sample.

[0150] Therefore, according to the embodiments of this application, basophils and nucleated red blood cells in the test sample can be accurately distinguished, and the information of infected red blood cells, white blood cell five-part differential and nucleated red blood cells in the test sample can be obtained through a single reaction and detection channel.

[0151] In some embodiments, the processor 140 is further configured to obtain nucleated red blood cell information of the test sample based at least on a third fluorescence signal. This enables the acquisition of infected red blood cell information, white blood cell differential results, and nucleated red blood cell information in a single test, significantly reducing detection costs.

[0152] In other embodiments, the processor 140 is further configured to: generate a first scatter plot based on at least one scattered light signal and a first fluorescence signal, and obtain infected red blood cell information based on the first scatter plot; generate a second scatter plot based on at least one scattered light signal and a second fluorescence signal, and generate a third scatter plot based on at least one scattered light signal and a third fluorescence signal; obtain white blood cell five-part differential results based on the second scatter plot and the third scatter plot; and obtain nucleated red blood cell information based on the third scatter plot.

[0153] In a specific example, obtaining the five-part differential white blood cell count based on the second and third scatter plots includes: obtaining the four-part differential white blood cell count of the test sample based on the second scatter plot, wherein the four-part differential white blood cell count includes the counts of lymphocytes, monocytes, neutrophils, and eosinophils in the test sample; identifying a first characteristic cell group including basophils and nucleated erythrocytes in the second scatter plot and obtaining the first cell count of the first characteristic cell group; identifying a second characteristic cell group including only nucleated erythrocytes in the third scatter plot and obtaining the second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the count of basophils in the test sample.

[0154] Here, the second cell count is the count of nucleated red blood cells in the test sample.

[0155] In the third embodiment of this application, the sampling device 110 is used to aspirate the blood sample to be tested; the sample preparation device 120 is used to mix at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample; the optical detection device 130 includes a light emitting device, a flow cell, and a photodetector. The light emitting device is used to emit excitation light to irradiate the flow cell, the flow cell is connected to the reaction cell and is used to allow individual particles in the test sample to pass through. The photodetector is used to detect the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow cell in a single test. The optical information includes at least one scattered light signal detected in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye. Here, the processor 140 is configured to obtain information on infected red blood cells and nucleated red blood cells of the test sample based on at least one scattered light signal, a first fluorescence signal, and a third fluorescence signal detected in a single test of the test sample prepared by mixing the blood sample to be tested, the hemolysin, the first fluorescent dye, and the third fluorescent dye.

[0156] In some embodiments, the first fluorescent dye may be a fluorescent dye that can be used to specifically label malaria parasites, and the third fluorescent dye may be a fluorescent dye that can be used to identify nucleated red blood cells.

[0157] In some embodiments, the processor 140 obtains information on infected red blood cells and nucleated red blood cells of the test sample based on at least one scattered light signal, a first fluorescence signal, and a third fluorescence signal, including: obtaining a count result of nucleated red blood cell populations in the test sample based on at least one scattered light signal and a third fluorescence signal; and obtaining a count result of infected red blood cells in the test sample and / or the type of Plasmodium infecting the infected red blood cells based on at least one scattered light signal and a first fluorescence signal.

[0158] In some embodiments, the processor 140 is further configured to obtain the white blood cell count result in the test sample based on at least one scattered light signal and a third fluorescence signal.

[0159] This application also proposes a sample analysis method 200, such as... Figure 3 As shown, it includes S210, S220, S230 and S240.

[0160] In step S210, a blood sample to be tested is drawn. For example, a portion of the blood sample to be tested is drawn from a test tube containing the blood sample to be tested.

[0161] In step S220, at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a second fluorescent dye are mixed to prepare a single test sample.

[0162] For example, in step S220, the hemolysin, the first fluorescent dye, and the second fluorescent dye can be added sequentially to the same blood sample to obtain a test sample. The test sample is then incubated to allow the dyes to fully stain the particles in the test sample. Alternatively, in step S220, the first and second fluorescent dyes can be pre-mixed with the hemolysin to obtain a mixed reagent. This mixed reagent is then mixed with the blood sample to be tested at a volume ratio of 250:1 to 1000:1. After thorough mixing, the resulting test sample is incubated at 25°C to 50°C for 10 seconds to 1 minute, preferably 10 seconds to 40 seconds.

[0163] Examples of the hemolytic agent and fluorescent dye used in step S220 can be found in the above description and will not be repeated here.

[0164] In step S230, each particle in the test sample is passed through an optical detection area irradiated by excitation light in a single test to obtain the optical information generated by each particle in the test sample after being irradiated by excitation light. The excitation light includes light of a first wavelength that can excite the first fluorescent dye and light of a second wavelength that can excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal obtained in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a second fluorescence signal corresponding to the second fluorescent dye.

[0165] Here, the first fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the first fluorescent dye under the excitation of light at the first wavelength, while the second fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the second fluorescent dye under the excitation of light at the second wavelength.

[0166] In other words, in step S230, the scattered light signal (or scattered light signal intensity) and fluorescence signal (or fluorescence signal intensity) of the sample are obtained based on the principle of flow cytometry.

[0167] In step S240, the infected red blood cell information of the test sample is obtained based on at least one scattered light signal and a first fluorescence signal; and the white blood cell four-part differential result of the test sample is obtained based on at least one scattered light signal and a second fluorescence signal, wherein the white blood cell four-part differential result includes the count results of lymphocyte population, monocyte population, neutrophil population and eosinophil population in the test sample.

[0168] According to another aspect of this application, a sample analysis method 300 is proposed, such as... Figure 4 As shown, it includes S310, S320, S330 and S340.

[0169] In step S310, a blood sample to be tested is drawn. For example, a portion of the blood sample to be tested is drawn from a test tube containing the blood sample to be tested.

[0170] In step S320, at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye are mixed to prepare a single test sample.

[0171] For example, in step S320, the hemolysin, the first fluorescent dye, the second fluorescent dye, and the third fluorescent dye can be added sequentially to the same blood sample to obtain a test sample. The test sample is then incubated to allow the dyes to fully stain the particles in the test sample. Alternatively, in step S320, the first, second, and third fluorescent dyes can be pre-mixed with the hemolysin to obtain a mixed reagent. This mixed reagent is then mixed with the blood sample to be tested at a volume ratio of 250:1 to 1000:1. After thorough mixing, the resulting test sample is incubated at 25°C to 50°C for 10 seconds to 1 minute, preferably 10 seconds to 40 seconds.

[0172] Examples of the hemolytic agent and fluorescent dye used in step S320 can be found in the above description and will not be repeated here.

[0173] In step S330, each particle in the test sample is passed through an optical detection area irradiated by excitation light in a single test to obtain the optical information generated by each particle in the test sample after being irradiated by excitation light. The excitation light includes light of a first wavelength that can excite the first fluorescent dye and light of a second wavelength that can excite the third fluorescent dye. The first wavelength light or the second wavelength light can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal obtained in a single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0174] Here, the first fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the first fluorescent dye under light excitation at the first wavelength, the second fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the second fluorescent dye under light excitation at the first wavelength, and the third fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the third fluorescent dye under light excitation at the second wavelength.

[0175] In other words, in step S330, the scattered light signal (or scattered light signal intensity) and fluorescence signal (or fluorescence signal intensity) of the sample are obtained based on the principle of flow cytometry.

[0176] In step S340, the infected red blood cell information and white blood cell five-part differential results of the test sample are obtained based on at least one scattered light signal, a first fluorescence signal, a second fluorescence signal and a third fluorescence signal. The white blood cell five-part differential results include the count results of lymphocyte population, monocyte population, neutrophil population, eosinophil population and basophil population in the test sample.

[0177] According to another aspect of this application, a sample analysis method 400 is proposed, such as... Figure 5 As shown, it includes S410, S420, S430, and S440. It includes:

[0178] In step S410, a blood sample to be tested is drawn. For example, a portion of the blood sample to be tested is drawn from a test tube containing the blood sample to be tested.

[0179] In step S420, at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a third fluorescent dye are mixed to prepare a single test sample.

[0180] For example, in step S420, the hemolysin, the first fluorescent dye, and the third fluorescent dye can be added sequentially to the same blood sample to obtain a test sample. The test sample is then incubated to allow the dyes to fully stain the particles in the test sample. Alternatively, in step S420, the first and third fluorescent dyes can be pre-mixed with the hemolysin to obtain a mixed reagent. This mixed reagent is then mixed with the blood sample to be tested at a volume ratio of 250:1 to 1000:1. After thorough mixing, the resulting test sample is incubated at 25°C to 50°C for 10 seconds to 1 minute, preferably 10 seconds to 40 seconds.

[0181] Examples of the hemolytic agent and fluorescent dye used in step S420 can be found in the above description and will not be repeated here.

[0182] In step S430, each particle in the test sample is passed through an optical detection area irradiated by excitation light in a single test to obtain optical information generated by each particle in the test sample after being irradiated by excitation light. The optical information includes at least one scattered light signal obtained in a single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye.

[0183] Here, the first fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the first fluorescent dye under the excitation of light at the first wavelength, and the third fluorescence signal is the fluorescence signal generated by the binding of particles in the sample with the third fluorescent dye under the excitation of light at the second wavelength.

[0184] In other words, in step S430, the scattered light signal (or scattered light signal intensity) and fluorescence signal (or fluorescence signal intensity) of the sample are obtained based on the principle of flow cytometry.

[0185] In step S440, the information on infected red blood cells and nucleated red blood cells of the test sample is obtained based on at least one scattered light signal, a first fluorescence signal, and a third fluorescence signal.

[0186] The present application is described below with reference to embodiments intended to illustrate the application and not limit it. Unless otherwise specified, the experiments and methods described in the embodiments are generally performed according to conventional methods well known in the art and described in various references. Furthermore, where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0187] Example 1

[0188] The reagent formula is as follows:

[0189] A: Fluorescent dye solution

[0190] 30mg of the first fluorescent dye

[0191] 50mg of the second fluorescent dye

[0192] 1000g of ethylene glycol

[0193] B: Hemolytic agent

[0194]

[0195] The first fluorescent dye has the following general formula:

[0196]

[0197] The second fluorescent dye has the following general formula:

[0198]

[0199] The sample analyzer according to this application, equipped with a red laser (emitting a beam with a wavelength of approximately 633 nm) and a blue laser (emitting a beam with a wavelength of approximately 488 nm), was used to process and detect uninfected and infected blood samples, respectively: 20 μL of reagent A and 20 μL of the blood sample were added to 1 mL of reagent B to form a test sample; the intensity of scattered light corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation were detected; based on the first fluorescence intensity and the intensity of scattered light (forward scattered light intensity or lateral scattered light intensity), the results were analyzed. A two-dimensional first scatter plot is generated based on the scattered light intensity. The region Infect_Area representing infected red blood cells is obtained from the first scatter plot. The scatter points falling into this region are counted to obtain the count value of infected red blood cells. A two-dimensional second scatter plot is generated based on the second fluorescence intensity and the side scattered light intensity. The white blood cell four-part differential result is obtained based on the second scatter plot. The white blood cell four-part differential result includes the count results of neutrophils (neu), lymphocytes (lym), monocytes (mon), and eosinophils (eos) in the sample.

[0200] Figure 6a and Figure 6b The first scatter plot shows the results of processing and testing uninfected blood samples using a sample analyzer. Figure 6c The second scatter plot is shown, obtained by processing and detecting a blood sample that was not infected with malaria parasites using a sample analyzer. In the first scatter plot in Figure 6, there are virtually no scatter points in the Infect_Area (region of infected red blood cells), indicating that the blood sample was not infected with malaria parasites.

[0201] Figure 7a and Figure 7b The first scatter plot shows the results of processing and testing a blood sample infected with malaria parasites using a sample analyzer. Figure 7c The second scatter plot is shown, obtained from the processing and detection of a blood sample infected with malaria parasites by a sample analyzer. The presence of a certain number of scatter points in the Infect_Area (the area of ​​infected red blood cells) of the first scatter plot in Figure 7 indicates that the blood sample was infected with malaria parasites.

[0202] Example 2

[0203] The sample analyzer according to this application, equipped with a red laser (emitting a beam with a wavelength of approximately 633 nm) and a blue laser (emitting a beam with a wavelength of approximately 488 nm), was used to process and detect blood samples infected with different types of Plasmodium parasites: 20 μL of the above-mentioned reagent A (reagent A of Example 1) and 20 μL of the blood sample to be tested were added to 1 mL of the above-mentioned reagent B (reagent B of Example 1) to form a test sample; the forward scattered light intensity corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation were detected; a two-dimensional first scatter plot was generated based on the first fluorescence intensity and the forward scattered light intensity; the region Infect_Area representing infected red blood cells was obtained from the first scatter plot; by identifying the range of the distribution peak of the forward scattered light intensity within this region, the type of Plasmodium parasite infecting the infected red blood cells could be determined.

[0204] Figure 8a The first scatter plot shows the results of processing and testing a blood sample infected with Plasmodium falciparum using a sample analyzer. Figure 8b The first scatter plot shows the results of processing and testing blood samples infected with Plasmodium ovale using a sample analyzer. Figure 8c The first scatter plot is shown, obtained by processing and detecting blood samples infected with Plasmodium vivax using a sample analyzer. Due to differences in the size of different types of Plasmodium, the intensity of the forward scattered light signal from erythrocytes infected with Plasmodium falciparum, Plasmodium ovale, and Plasmodium vivax differs. Specifically, the centroid of the forward scattered light intensity of erythrocytes infected with Plasmodium vivax is larger than that of erythrocytes infected with Plasmodium ovale, and the centroid of the forward scattered light intensity of erythrocytes infected with Plasmodium ovale is larger than that of erythrocytes infected with Plasmodium falciparum.

[0205] Example 3

[0206] The sample analyzer according to this application, equipped with a red laser (emitting a beam with a wavelength of approximately 633 nm) and a blue laser (emitting a beam with a wavelength of approximately 488 nm), was used to process and detect uninfected and infected blood samples. 20 μL of reagent A (reagent A of Example 1) and 20 μL of the blood sample were added to 1 mL of reagent B (reagent B in Example 1) to form a test sample. The intensity of scattered light corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation were detected.

[0207] A two-dimensional first scatter plot is generated based on the first fluorescence intensity and the scattered light intensity (forward scattered light intensity or side scattered light intensity), as follows: Figure 9a and Figure 9b As shown, based on the first scatter plot, the region Infect_Area representing infected red blood cells is obtained, and the scatter points falling into this region are counted to obtain the count value of infected red blood cells; and a two-dimensional second scatter plot is generated based on the second fluorescence intensity and the side-scatter light intensity, as shown. Figure 9c As shown, the white blood cell four-part differential results are obtained based on the second scatter plot. The white blood cell four-part differential results include the count of neutrophils (neu), lymphocytes (lym), monocytes (mon), and eosinophils (eos) in the sample.

[0208] Here, based on Figure 9a and Figure 9b The first scatter plot shown is divided into regions representing infected red blood cells, Infect_Area. From this region Infect_Area, the first infected red blood cell group Infect_CellPop1 and the second infected red blood cell group Infect_CellPop2 are identified. Each infected red blood cell in the second infected red blood cell group Infect_CellPop2 contains multiple malaria parasite rings.

[0209] Example 4

[0210] The reagent formula is as follows:

[0211] A: Fluorescent dye solution

[0212]

[0213] B: Hemolytic agent

[0214]

[0215] The first fluorescent dye has the following general formula:

[0216]

[0217] The second fluorescent dye has the following general formula:

[0218]

[0219] The third fluorescent dye has the following general formula:

[0220]

[0221] The sample analyzer according to this application, equipped with a red laser (emitting a beam with a wavelength of approximately 633 nm) and a blue laser (emitting a beam with a wavelength of approximately 488 nm), was used to process and detect blood samples infected with Plasmodium: 20 μL of reagent A and 20 μL of the blood sample to be tested were added to 1 mL of reagent B to form a test sample; the forward and side-scattered light intensities corresponding to blue light excitation, the first and second fluorescence intensities corresponding to blue light excitation, and the third fluorescence intensity corresponding to red light excitation were detected.

[0222] A two-dimensional first scatter plot is generated based on the first fluorescence intensity and the scattered light intensity (forward scattered light intensity or side scattered light intensity), as follows: Figure 10a and Figure 10b As shown, based on the first scatter plot, the region Infect_Area representing infected red blood cells is obtained. Scatter points falling into this region are counted to obtain the count value of infected red blood cells. A two-dimensional second scatter plot is generated based on the second fluorescence intensity and the side-scatter light intensity, as shown below. Figure 10c As shown; and a two-dimensional third scatter plot generated based on the third fluorescence intensity and the side-scattered light intensity, as shown. Figure 10d As shown. Here, the five-part differential white blood cell count is obtained based on the second and third scatter plots. This five-part differential white blood cell count includes the counts of neutrophils (neu), lymphocytes (lym), monocytes (mon), eosinophils (eos), and basophils (baso) in the sample. The count of nucleated red blood cells (NRBC) is obtained based on the third scatter plot.

[0223] Example 5

[0224] The reagent formula is as follows:

[0225] A: Fluorescent dye solution

[0226] 30mg of the first fluorescent dye

[0227] 30mg of the third fluorescent dye

[0228] 1000g of ethylene glycol

[0229] B: Hemolytic agent

[0230]

[0231] The first fluorescent dye has the following general formula:

[0232]

[0233] The third fluorescent dye has the following general formula:

[0234]

[0235] The sample analyzer according to this application, equipped with a red laser (emitting a beam with a wavelength of approximately 633 nm) and a blue laser (emitting a beam with a wavelength of approximately 488 nm), was used to process and detect blood samples infected with Plasmodium: 20 μL of reagent A and 20 μL of the blood sample to be tested were added to 1 mL of reagent B to form a test sample; the forward and side-scattered light intensities corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation were detected.

[0236] A two-dimensional scatter plot is generated based on the first fluorescence intensity and the forward scattered light intensity, as follows: Figure 11a As shown, based on the first scatter plot, the region Infect_Area representing infected red blood cells is obtained. Scatter points falling into this region are counted to obtain the count value of infected red blood cells. A two-dimensional third scatter plot is generated based on the second fluorescence intensity and the side-scatter light intensity, as shown below. Figure 11b As shown, the count of nucleated erythrocytes (NRBC) was obtained based on the third scatter plot.

[0237] 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 with reference to the sample analyzer provided in this application shall be applied accordingly to the sample analysis method provided in this application, and vice versa.

[0238] The above description is merely a preferred embodiment of this application and does not limit the scope of patent protection of this application. All equivalent modifications made based on the content of the specification and drawings of this application, or direct / indirect applications in other related technical fields, are included within the scope of patent protection of this application.

Claims

1. A sample analyzer, comprising: A sampling device is used to collect blood samples for testing. A sample preparation apparatus for mixing at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a second fluorescent dye in a reaction cell to prepare a test sample; An optical detection device includes a light emitting device, a flow chamber, and a photodetector. The light emitting device emits excitation light to irradiate the flow chamber, which is connected to a reaction cell and allows individual particles in the test sample to pass through. The photodetector detects the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow chamber in a single test. The light emitting device includes a first light source and a second light source. The first light source emits light of a first wavelength capable of exciting a first fluorescent dye, and the second light source emits light of a second wavelength capable of exciting a second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in the single test, a first fluorescence signal corresponding to the first fluorescent dye, and a second fluorescence signal corresponding to the second fluorescent dye; and The processor is configured to: obtain infected red blood cell information of the test sample based on at least one of the scattered light signals and the first fluorescence signal; and obtain a four-part differential white blood cell count of the test sample based on at least one of the scattered light signals and the second fluorescence signal, wherein the four-part differential white blood cell count includes the count of lymphocytes, the count of monocytes, the count of neutrophils and the count of eosinophils in the test sample.

2. The sample analyzer according to claim 1, characterized in that, The first wavelength is between 315nm and 490nm and the second wavelength is between 610nm and 750nm.

3. The sample analyzer according to claim 1 or 2, characterized in that, The scattered light signal includes: a first forward scattered light signal and a first side scattered light signal generated by the particles passing through the flow chamber after being irradiated by light of the first wavelength, as detected in the single test; and / or a second forward scattered light signal and a second side scattered light signal generated by the particles passing through the flow chamber after being irradiated by light of the second wavelength, as detected in the single test. The processor obtains the infected erythrocyte information of the test sample based on at least one of the scattered light signals and the first fluorescence signal, including: A first scatter plot is generated based on one of the first forward scattered light signal, the first side scattered light signal, the second forward scattered light signal, the second side scattered light signal, and the first fluorescence signal, and the infected red blood cell information of the test sample is obtained based on the first scatter plot.

4. The sample analyzer according to claim 3, characterized in that, The processor obtains the infected red blood cell information of the test sample based on the first scatter plot, including: The count of infected red blood cells in the test sample is obtained based on the first scatter plot; and / or The type of malaria parasite infecting the infected red blood cells in the test sample is determined based on the first scatter plot.

5. The sample analyzer according to claim 4, characterized in that, The processor generates a first scatter plot based on one of the first forward-scattered light signal, the first side-scattered light signal, the second forward-scattered light signal, the second side-scattered light signal, and the first fluorescence signal, including: generating the first scatter plot based on the first forward-scattered light signal and the first fluorescence signal or based on the second forward-scattered light signal and the first fluorescence signal; The processor determines the type of malaria parasite infected by the infected red blood cells in the test sample based on the first scatter plot, including: identifying infected red blood cell regions containing infected red blood cell clusters from the first scatter plot, and determining whether the malaria parasite infected by the infected red blood cell clusters in the test sample is Plasmodium falciparum, Plasmodium ovale, or Plasmodium vivax based on the forward scattered light intensity of the infected red blood cell clusters in the infected red blood cell regions.

6. The sample analyzer according to any one of claims 1 to 5, characterized in that, The scattered light signal includes: a first side-scattered light signal generated by a particle passing through the flow chamber after being irradiated by light of the first wavelength, detected in the single test, and / or a second side-scattered light signal generated by a particle passing through the flow chamber after being irradiated by light of the second wavelength, detected in the single test. The processor obtains the white blood cell four-part differential result of the test sample based on at least one of the scattered light signals and the second fluorescence signal, including: A second scatter plot is generated based on the first side-scattered light signal and the second fluorescence signal, or based on the second side-scattered light signal and the second fluorescence signal, and the white blood cell four-part differential result of the test sample is obtained based on the second scatter plot.

7. A sample analyzer, comprising: A sampling device is used to collect blood samples for testing. A sample preparation apparatus for mixing at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample; An optical detection device includes a light emitting device, a flow chamber, and a photodetector. The light emitting device emits excitation light to irradiate the flow chamber, which is connected to a reaction cell and allows particles in the test sample to pass through. The photodetector detects the optical information generated when particles in the test sample are irradiated by the excitation light as they pass through the flow chamber in a single test. The light emitting device includes a first light source and a second light source. The first light source emits light capable of exciting a first fluorescent dye at a first wavelength, and the second light source emits light capable of exciting a third fluorescent dye at a second wavelength. Either the first or second wavelength light can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal detected in the single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye. The processor is configured to obtain information on infected red blood cells and white blood cell differential counts of the test sample based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal, and the third fluorescence signal, wherein the white blood cell differential counts include the counts of lymphocytes, monocytes, neutrophils, eosinophils, and basophils in the test sample.

8. The sample analyzer according to claim 7, characterized in that, The processor obtains the infected red blood cell information and white blood cell five-part differential results of the test sample based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal, and the third fluorescence signal, including: The infected erythrocyte information is obtained based on at least one of the scattered light signal, the second fluorescence signal, the third fluorescence signal, and the first fluorescence signal; and The five-part differential diagnosis of white blood cells is obtained based on at least one of the scattered light signals, the second fluorescence signal, and the third fluorescence signal.

9. The sample analyzer according to claim 8, characterized in that, The processor obtains the infected red blood cell information based on at least one of the scattered light signal, the second fluorescence signal, the third fluorescence signal, and the first fluorescence signal, including: A first scatter plot is generated based on at least one of the scattered light signals and the first fluorescence signal, and the infected red blood cell information is obtained based on the first scatter plot; and The processor obtains the five-part differential diagnosis of white blood cells based on at least one of the scattered light signals, the second fluorescence signal, and the third fluorescence signal, including: A second scatter plot is generated based on at least one of the scattered light signals and the second fluorescence signal. The white blood cell differential count of the test sample was obtained based on the second scatter plot, wherein the white blood cell differential count included the counts of lymphocytes, monocytes, neutrophils, and eosinophils in the test sample. The first characteristic cell group, including basophils, is delineated in the second scatter plot, and the count of basophils in the test sample is obtained based on at least a third fluorescence signal of the first characteristic cell group.

10. The sample analyzer according to claim 8, characterized in that, The processor obtains the infected red blood cell information based on at least one of the scattered light signal, the second fluorescence signal, the third fluorescence signal, and the first fluorescence signal, including: A first scatter plot is generated based on at least one of the scattered light signals and the first fluorescence signal, and the infected red blood cell information is obtained based on the first scatter plot; and The processor obtains the five-part differential diagnosis of white blood cells based on at least one of the scattered light signals, the second fluorescence signal, and the third fluorescence signal, including: A second scatter plot is generated based on at least one of the scattered light signals and the second fluorescence signal, and a four-part differential white blood cell count of the test sample is obtained based on the second scatter plot, wherein the four-part differential white blood cell count includes the counts of lymphocytes, monocytes, neutrophils, and eosinophils in the test sample; and A third scatter plot is generated based on at least one of the scattered light signals and the third fluorescence signal, and the count of basophils in the test sample is obtained based on the second scatter plot and the third scatter plot.

11. The sample analyzer according to claim 10, characterized in that, The processor obtains the basophil count results in the test sample based on the second scatter plot and the third scatter plot, including: In the second scatter plot, a first characteristic cell group including basophils and nucleated erythrocytes is identified and a first cell count of the first characteristic cell group is obtained. The second characteristic cell group, consisting only of nucleated erythrocytes, is segmented in the third scatter plot, and a second cell count of this second characteristic cell group is obtained; and The second cell count is subtracted from the first cell count to obtain the basophil population count of the test sample.

12. The sample analyzer according to claim 7 or 8, characterized in that, The processor is also configured to obtain nucleated red blood cell information of the test sample based at least on the third fluorescence signal.

13. The sample analyzer according to claim 12, characterized in that, The processor is also configured to: A first scatter plot is generated based on at least one of the scattered light signals and the first fluorescence signal, and the information of the infected red blood cells is obtained based on the first scatter plot; A second scatter plot is generated based on at least one of the scattered light signals and the second fluorescence signal, and a third scatter plot is generated based on at least one of the scattered light signals and the third fluorescence signal; The white blood cell five-part differential result is obtained based on the second scatter plot and the third scatter plot; as well as The nucleated red blood cell information is obtained based on the third scatter plot.

14. The sample analyzer according to claim 13, characterized in that, The process of obtaining the five-part differential diagnosis of white blood cells based on the second scatter plot and the third scatter plot includes: The white blood cell four-part differential result of the test sample is obtained based on the second scatter plot, wherein the white blood cell four-part differential result includes the count results of lymphocytes, monocytes, neutrophils and eosinophils in the test sample. In the second scatter plot, a first characteristic cell group including basophils and nucleated erythrocytes is identified and a first cell count of the first characteristic cell group is obtained. The second characteristic cell group, consisting only of nucleated erythrocytes, is segmented in the third scatter plot, and a second cell count of this second characteristic cell group is obtained; and The second cell count is subtracted from the first cell count to obtain the basophil population count of the test sample.

15. A sample analyzer, comprising: A sampling device is used to collect blood samples for testing. A sample preparation apparatus for mixing at least a portion of the blood sample to be tested, a hemolysin, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample; An optical detection device includes a light emitting device, a flow chamber, and a photodetector. The light emitting device is used to emit excitation light to irradiate the flow chamber. The flow chamber is connected to the reaction cell and is used to allow individual particles in the test sample to pass through. The photodetector is used to detect the optical information generated when individual particles in the test sample are irradiated by the excitation light as they pass through the flow chamber in a single test. The optical information includes at least one scattered light signal detected in the single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye. as well as The processor is configured to obtain information on infected red blood cells and nucleated red blood cells of the test sample based on at least one of the scattered light signals, the first fluorescence signal, and the third fluorescence signal.

16. The sample analyzer according to claim 15, characterized in that, The light emitting device includes a first light source and a second light source. The first light source is used to emit light of a first wavelength that can excite the first fluorescent dye, and the second light source is used to emit light of a second wavelength that can excite the third fluorescent dye. The second wavelength is greater than the first wavelength. Preferably, the first wavelength is between 315 nm and 490 nm and the second wavelength is between 610 nm and 750 nm. The processor obtains the infected erythrocyte information and nucleated erythrocyte information of the test sample based on at least one of the scattered light signals, the first fluorescence signal, and the third fluorescence signal, including: The count of nucleated erythrocytes in the test sample is obtained based on at least one of the scattered light signals and the third fluorescence signal; and The count of infected red blood cells in the test sample and / or the type of malaria parasite infected by the infected red blood cells are obtained based on at least one of the scattered light signals and the first fluorescence signal.

17. The sample analyzer according to claim 15 or 16, characterized in that, The processor is also configured to: The white blood cell count in the test sample is obtained based on at least one of the scattered light signals and the third fluorescence signal.

18. A sample analysis method, comprising: Collect the blood sample to be tested; At least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye are mixed to prepare the test sample; In a single test, each particle in the test sample is passed through an optical detection area irradiated by excitation light to obtain optical information generated by each particle in the test sample after being irradiated by the excitation light. The excitation light includes light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal obtained in the single test, a first fluorescence signal corresponding to the first fluorescent dye, and a second fluorescence signal corresponding to the second fluorescent dye. The infected erythrocyte information of the test sample is obtained based on at least one of the scattered light signals and the first fluorescence signal; and The white blood cell four-part differential result of the test sample is obtained based on at least one of the scattered light signals and the second fluorescence signal, wherein the white blood cell four-part differential result includes the count results of lymphocytes, monocytes, neutrophils and eosinophils in the test sample.

19. A sample analysis method, comprising: Collect the blood sample to be tested; The test sample is prepared by mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye. In a single test, each particle in the test sample is passed through an optical detection area irradiated by excitation light to obtain optical information generated by each particle after being irradiated by the excitation light. The excitation light includes light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the third fluorescent dye. Either the first wavelength or the second wavelength can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one scattered light signal obtained in the single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye; and The infected red blood cell information and white blood cell five-part differential results of the test sample are obtained based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, wherein the white blood cell five-part differential results include the count results of lymphocyte population, monocyte population, neutrophil population, eosinophil population and basophil population in the test sample.

20. A sample analysis method, comprising: Collect the blood sample to be tested; The test sample is prepared by mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye. In a single test, each particle in the test sample is passed through an optical detection area irradiated by excitation light to obtain optical information generated by each particle in the test sample after being irradiated by the excitation light. The optical information includes at least one scattered light signal obtained in the single test, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye. as well as The information on infected red blood cells and nucleated red blood cells of the test sample is obtained based on at least one of the scattered light signals, the first fluorescence signal, and the third fluorescence signal.

Images