WO2023207906A1 - 检测原始细胞的血细胞分析仪及检测方法 - Google Patents

检测原始细胞的血细胞分析仪及检测方法 Download PDF

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Publication number
WO2023207906A1
WO2023207906A1 PCT/CN2023/090310 CN2023090310W WO2023207906A1 WO 2023207906 A1 WO2023207906 A1 WO 2023207906A1 CN 2023090310 W CN2023090310 W CN 2023090310W WO 2023207906 A1 WO2023207906 A1 WO 2023207906A1
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Prior art keywords
sample
reagent
information
signal
detection
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English (en)
French (fr)
Inventor
李进
秦哲原
陈庚文
姚栋蓝
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Shenzhen Mindray Bio Medical Electronics Co Ltd
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Shenzhen Mindray Bio Medical Electronics Co Ltd
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Priority to CN202380035463.9A priority Critical patent/CN119072621A/zh
Priority to EP23795339.3A priority patent/EP4517304A4/en
Publication of WO2023207906A1 publication Critical patent/WO2023207906A1/zh
Priority to US18/927,686 priority patent/US20250052680A1/en
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • G01N2015/016—White blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1486—Counting the particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks

Definitions

  • the present disclosure relates to the field of in vitro diagnostics, and in particular to blood cell analyzers and methods for primary cell detection.
  • Explosive cells are not usually present in the peripheral blood of healthy individuals.
  • the presence of blast cells in peripheral blood usually indicates hematological diseases, such as acute and chronic leukemia, myelodysplastic syndrome, etc.
  • hematological diseases such as acute and chronic leukemia, myelodysplastic syndrome, etc.
  • acute leukemia is divided into two categories: acute lymphoblastic leukemia and acute myeloid leukemia and their different subtypes.
  • MICM morphology, immunology, cytogenetics, molecular biology
  • peripheral blood examination can be used as a means to evaluate the clinical diagnosis and treatment of AML.
  • the clearance rate of blast cells in the patient's peripheral blood after induction chemotherapy and the level of residual leukemia cells in the peripheral blood after chemotherapy are closely related to the patient's sensitivity and efficacy to the drug.
  • the ratio of blast cells in peripheral blood to initial blast cells in the peripheral blood can be continuously monitored for 1 to 7 days. The number of days required for the ratio to reach 0.1 is an independent prognostic factor for AML.
  • the hematology analyzer method is currently used for screening in clinical laboratories and is based on the blast cell alarm rules of the International Hematology Reexamination Expert Group, triggering manual microscopy to screen and confirm the presence of blast cells. Therefore, although the blood cell analyzer is fast, efficient, and easy to operate, it cannot accurately identify and quantify original cells and can only give suspicious alarms. This alarm has a high false negative rate and a high false positive rate. Positive rate.
  • Microscopy relies on manual or software observation of blood smears and is an industry-recognized reference method for primary cell classification and counting.
  • the number of cells detected by this method is small, only 100-200 per person. Therefore, the quality and level of the detection results are affected by specimen collection, smears, staining, etc., and it is difficult to detect precision and sensitivity of original cells. Significant improvement.
  • the flow cytometry analyzer method uses flow cytometry technology and uses a variety of monoclonal antibodies to classify and count leukocytes and other blood cells based on differences in the expression of antigens on the surface of leukocytes and other blood cells. Therefore, this method eliminates the subjective errors introduced by manual microscopy and significantly increases the statistical volume. It is a method that is expected to become more accurate and efficient. However, this method is expensive and the operation is cumbersome and complicated.
  • the International Committee for Standardization in Hematology (ICSH) organized five reference laboratories around 2016 to explore flow cytometry methods to classify and count various subtypes of white blood cells, immature granulocytes, blast cells and nucleated red blood cells in peripheral blood. , eight monoclonal antibodies need to be used simultaneously. Since the cell surface antigens of primitive cells are non-specific, their detection relies on the elimination method.
  • the first object of the present disclosure is to provide a blood cell analyzer for primary cell detection and a primary cell
  • the detection method, the blood cell analyzer and the method can quickly and accurately analyze primitive cells in peripheral blood and can count them without affecting the classification and counting of other types of cells in peripheral blood samples.
  • a first aspect of the present disclosure provides a blood analyzer, including a sample suction device, a reagent supply device, a mixing chamber, a light source, an optical flow chamber, an optical detector, a processor and a non-transitory computer programmed with a computer application program readable storage medium, where,
  • the sample suction device is used to suck a sample and transport at least part of the sucked sample to the mixing chamber as a first sub-sample;
  • the reagent supply device is used to provide reagents and transport them to the mixing chamber, where the reagents include first type reagents;
  • the mixing chamber is used to mix the first sub-sample with the first type of reagent to form a first test sample; wherein the first type of reagent includes a first reagent and a second reagent, and the first The reagent includes a first hemolytic agent, the second reagent includes a fluorescently labeled antibody capable of binding to a surface antigen of the original cell;
  • the light source is used to align the light beam with the detection hole of the optical flow chamber connected to the mixing chamber;
  • the optical detector is used to detect the light scattering signal and the fluorescence signal of the first test sample passing through the detection hole.
  • the light scattering signal includes the first lateral light scattering intensity from the first test sample.
  • a signal, the fluorescent signal comprising an antibody fluorescence intensity signal from the fluorescently labeled antibody;
  • the processor is operatively connected to the optical detector, and when the computer application program in the non-transitory computer-readable storage medium is executed by the processor, the first hemolytic agent and the third hemolytic agent are controlled to The mixing time of a subsample in the mixing chamber does not exceed 2 minutes, and the processor obtains the particles in the first subsample based on the first side light scattering intensity signal and the antibody fluorescence intensity signal. Information, the particle information in the first subsample includes original cell information.
  • the first type of reagent further includes a third reagent
  • the third reagent includes a first fluorescent dye
  • the light scattering signal further includes a first fluorescence signal from the first fluorescent dye
  • the processor further obtains particle information in the first subsample based on the first side light scattering intensity signal and the first fluorescence intensity signal, so The particle information in the first subsample includes neutrophil information, monocyte information, lymphocyte information and eosinophil information.
  • the processor when the computer application is executed by the processor, the processor further: when the original cell information shows that the sample does not contain original cells, according to the first sub-sample Particle information, obtain classified counts of neutrophils, eosinophils, monocytes and lymphocytes, and optionally obtain immature granulocyte information and/or atypical lymphocyte information; or, the processor is configured according to The obtained original cell information, when the original cell information shows that the sample contains original cells, deducts the influence of the original cells from the particle information of the first sub-sample to obtain neutrophils, eosinophils Differential counting of granulocytes, monocytes and lymphocytes, and optionally obtaining immature granulocyte information and/or atypical lymphocyte information.
  • the sample suction device also sequentially delivers at least part of the suction sample to the mixing chamber as a second sub-sample and a third sub-sample;
  • the reagents also include a second type of reagent, and the The second type of reagents includes third reagents, fourth reagents, fifth reagents and sixth reagents;
  • the mixing chamber is further used to mix the second and third sub-samples respectively, wherein the second sub-sample is mixed with the third reagent and the fourth reagent to obtain the second test sample, and the third sub-sample is mixed with the third reagent.
  • the fifth reagent and the sixth reagent are mixed to obtain a third test sample; wherein the third reagent includes a first fluorescent dye, the fourth reagent includes a second hemolytic reagent, and the fifth reagent includes a second fluorescent dye , the sixth reagent includes a third hemolytic agent, and the second hemolytic agent is the same as or different from the first hemolytic agent, and the third hemolytic agent is different from both the first and second hemolytic agents, so The second fluorescent dye is different from the first fluorescent dye;
  • the optical detector is further used to detect the light scattering signal and the fluorescence signal of the second and third test samples respectively passing through the detection hole;
  • the light scattering signal includes a third signal from the second test sample.
  • the fluorescence signal includes a second fluorescence intensity signal from the first fluorescent dye of the second test sample and Second fluorescent dye from third test sample the third fluorescence intensity signal;
  • the processor When the computer application program is executed by the processor, the processor further obtains the second side light scattering intensity signal and the second fluorescence intensity signal of the second test sample. Particle information in the second subsample, and obtaining particle information in the third subsample based on the third side light scattering intensity signal and the third fluorescence intensity signal of the third test sample;
  • the particle information in the second subsample includes neutrophil information, monocyte information, lymphocyte information and eosinophil information
  • the particle information in the third subsample includes lymphocyte information, basophil information.
  • the processor when the computer application program is executed by the processor, the processor further: when the original cell information shows that the sample does not contain original cells, according to the particle information in the second sub-sample, Obtain differential counts of neutrophils, eosinophils, monocytes, and lymphocytes, and optionally immature granulocyte information and atypical lymphocyte information; or
  • the lymphocyte information in the original cell information and the particle information in the third subsample are combined, Eliminate the influence of the original cell information on the particle information in the second subsample, obtain differential counts of neutrophils, eosinophils, monocytes and lymphocytes, and optionally obtain immature granulocytes information and atypical lymphocyte information.
  • the third reagent may be included in the first type of reagents or in the second type of reagents.
  • the computer application program when executed by the processor, causes the processor to count primitive cells.
  • the processor when the computer application is executed by the processor, the processor further: when the original cell information shows that the sample contains original cells, an alarm is issued that there are original cells in the sample. , or alarm that there are primitive cells in the sample and output the primitive cell count value.
  • the light scattering signal further includes a first forward scattered light signal from the first test sample, and when the computer application program is executed by the processor, the processor
  • the primitive cells are further determined to be lymphoid primitive cells and/or myeloid primitive cells based on the first forward light scattering intensity signal and the antibody fluorescence intensity signal.
  • the light scattering signal further includes a first forward scattered light signal from the first test sample, and when the computer application program is executed by the processor, the processing The device further reports the presence of lymphoid blast cells and/or myeloid blast cells in the sample based on the first forward light scattering intensity signal and the antibody fluorescence intensity signal.
  • the processor when the computer application program is executed by the processor, the processor further obtains the concentration in the sample based on the first side light scattering intensity signal and the antibody fluorescence intensity signal.
  • the white blood cell information includes information about lymphocytes, monocytes, neutrophils and eosinophils.
  • the blood analyzer further includes a hemoglobin detection unit and/or an impedance counting detection unit.
  • a second aspect of the present disclosure provides a sample analysis method.
  • the method includes receiving operation instructions and performing original cell detection on the sample.
  • the original cell detection includes the following steps:
  • aspirating a sample including aspirating at least a portion of said sample as a first subsample
  • Processing the sample includes mixing the first sub-sample with a first type of reagent to obtain a first test sample; wherein the first type of reagent includes a first reagent and a second reagent, and the first reagent includes a first Hemolytic reagent, the mixing time of the first hemolytic reagent and the first sub-sample does not exceed 2 minutes, the second reagent includes a fluorescently labeled antibody, the antibody can bind to the surface antigen of the original cell;
  • Detecting the sample to be tested includes causing the particles in the first sample to be tested to pass through the detection area of the optical detection device one by one, And use the light source of the optical detection device to irradiate the particles in the first test sample to measure the light scattering signal and fluorescence signal of the first test sample, where the light scattering signal includes the light scattering signal from the first test sample.
  • a first side light scatter intensity signal of the test sample, the fluorescence signal comprising an antibody fluorescence intensity signal from the fluorescently labeled antibody;
  • Obtaining particle information includes obtaining particle information in the first subsample according to the first side light scattering intensity signal and the antibody fluorescence intensity signal, where the particle information in the first subsample includes original cell information.
  • the first type of reagent further includes a third reagent
  • the third reagent includes a first fluorescent dye
  • the light scattering signal further includes a third reagent from the first fluorescent dye in the first test sample.
  • a fluorescence signal the original cell detection further includes: obtaining particle information in the first subsample according to the first side light scattering intensity signal and the first fluorescence intensity signal, the first subsample
  • the particle information in includes neutrophil information, monocyte information, lymphocyte information and eosinophil information.
  • the primary cell detection further includes:
  • the original cell information shows that the sample does not contain original cells
  • obtain classified counts of neutrophils, eosinophils, monocytes and lymphocytes based on the particle information in the first sub-sample, And optionally obtain immature granulocyte information and/or atypical lymphocyte information; or, when the original cell information shows that the sample contains original cells, deduct all the information from the particle information in the first sub-sample.
  • the influence of blast cells is described, and differential counts of neutrophils, eosinophils, monocytes and lymphocytes are obtained, and optionally immature granulocyte information and/or atypical lymphocyte information is obtained.
  • the aspirating the sample further includes aspirating at least part of the sample as a second sub-sample and a third sub-sample;
  • the processing the sample further includes treating the second and third sub-samples with a second type of reagent, wherein the second type of reagent includes a third reagent, a fourth reagent, a fifth reagent and a sixth reagent, and the second type of reagent is The two sub-samples are mixed with the third reagent and the fourth reagent to obtain the second test sample, and the third sub-sample is mixed with the fifth reagent and the sixth reagent to obtain the third test sample; wherein, The third reagent includes a first fluorescent dye, the fourth reagent includes a second hemolytic agent, the fifth reagent includes a second fluorescent dye, the sixth reagent includes a third hemolytic agent, and the second hemolytic agent The third hemolytic agent is the same as or different from the first hemolytic agent, the third hemolytic agent is different from both the first and second hemolytic agents, and the second fluorescent dye is different from the first fluorescent dye;
  • the detection of the sample to be tested further includes causing the particles in the second and third samples to be tested to pass through the detection area of the optical detection device one by one, and using the light source of the optical detection device to detect the second and third samples respectively.
  • the particles in the third sample to be tested are irradiated to measure the light scattering signals and fluorescence signals of the second and third samples to be tested; wherein the light scattering signal includes the second light scattering signal from the second sample to be tested.
  • the fluorescence signal includes the second fluorescence intensity signal from the first fluorescent dye in the second test sample and the second fluorescence intensity signal from the second test sample.
  • the third fluorescence intensity signal of the second fluorescent dye in the third sample to be tested is irradiated to measure the light scattering signals and fluorescence signals of the second and third samples to be tested; wherein the light scattering signal includes the second light scattering signal from the second sample to be tested.
  • the obtaining particle information further includes obtaining the particle information in the second sub-sample according to the second side light scattering intensity signal and the second fluorescence intensity signal of the second test sample, and according to the The third side light scattering intensity signal and the third fluorescence intensity signal of the third test sample are used to obtain the particle information in the third sub-sample; wherein the particle information in the second sub-sample includes Neutrophil information, monocyte information, lymphocyte information and eosinophil information, the particle information in the third subsample includes lymphocyte information, basophil information and nucleated red blood cell information; and,
  • the original cell detection further includes:
  • the original cell information shows that the sample does not contain original cells
  • the lymphocyte information in the original cell information and the particle information in the third subsample are combined, Eliminate the influence of the original cell information on the particle information in the second subsample, and obtain neutrophils, eosinophils Differential counts of cells, monocytes, and lymphocytes, and optionally information on immature granulocytes and atypical lymphocytes.
  • the method further includes subjecting the sample to routine testing.
  • routine detection includes the following steps:
  • Processing the first initial sub-sample includes mixing the first initial sub-sample with a third reagent and a fourth reagent to obtain a first initial test sample, wherein the third reagent includes a first fluorescent dye, so The fourth reagent includes a second hemolytic agent, and the second hemolytic agent is the same as or different from the first hemolytic agent;
  • the particles in the first initial test sample are allowed to pass through the detection area of the optical detection device one by one and the light source of the optical detection device is used to illuminate the particles in the first initial test sample to obtain the The light scattering signal and the fluorescence signal of the particles in the first initial test sample, the light scattering signal includes the first initial side light scattering intensity signal from the first initial test sample; the fluorescence signal includes the first initial side light scattering intensity signal from the first test sample.
  • routine detection includes the following steps:
  • Processing the first and second initial sub-samples separately includes mixing the first initial sub-sample with a third reagent and a fourth reagent to obtain a first initial test sample, and mixing the second initial sub-sample Mix with a fifth reagent and a sixth reagent to obtain a second initial test sample.
  • the third reagent includes a first fluorescent dye.
  • the fourth reagent includes a second hemolytic reagent.
  • the fifth reagent includes a second fluorescent dye.
  • the sixth reagent includes a third hemolytic agent, and the second hemolytic agent is the same as or different from the first hemolytic agent, and the third hemolytic agent is different from both the first and second hemolytic agents, and the second fluorescent dye is different from the first fluorescent dye;
  • the particles in the first and second initial samples to be tested are respectively made to pass through the detection area of the optical detection device one by one, and the light source of the optical detection device is used to detect the particles in the first and second initial samples to be tested respectively.
  • the particles are irradiated to obtain light scattering signals and fluorescence signals of the particles in the first and second initial test samples, and the light scattering signals include the first initial lateral light from the first initial test sample.
  • the fluorescence signal includes the first initial fluorescence signal from the first fluorescent dye in the first initial test sample and the first initial fluorescence signal from the first test sample. 2.
  • the particle information of the first initial test sample is obtained according to the first initial side light scattering intensity signal and the first initial fluorescence intensity signal.
  • the particle information at least includes initial white blood cell information, and according to the second initial The side light scattering intensity signal and the second initial fluorescence intensity signal are used to obtain the particle information of the second initial test sample, and the particle information at least includes initial lymph information;
  • performing the above-mentioned primary cell detection on the sample further includes:
  • the routine detection is performed before the original detection, and further the operation instruction can be issued based on the results of the routine detection.
  • the operation instructions include instructions input by the user or instructions automatically triggered when the original cell alarm is issued.
  • the method further includes counting the original cells.
  • the method further includes: when the original cell information shows that the sample contains When primitive cells are detected, an alarm is issued that primitive cells are present in the sample, or a primitive cell count value is output.
  • the light scattering signal further includes a first forward scattered light signal from the first test sample
  • the method further includes: based on the first forward light scattering intensity signal and the The antibody fluorescence intensity signal determines that the primitive cells are lymphoid primitive cells and/or myeloid primitive cells.
  • the light scattering signal further includes a first forward scattered light signal from the first test sample
  • the method further includes: according to the first forward light scattering intensity signal and The antibody fluorescence intensity signal alerts to the presence of lymphoid blast cells and/or myeloid blast cells in the sample.
  • the method further includes: further obtaining leukocyte information in the sample according to the first side light scattering intensity signal and the antibody fluorescence intensity signal, where the leukocyte information includes lymphocytes, monocytes, Neutrophil and eosinophil information
  • the interaction time between the first hemolytic agent and the sample in the present disclosure is 30 to 80 seconds, preferably 50 to 60 seconds.
  • the first hemolytic agent described in this disclosure is capable of completely disrupting red blood cells while maintaining intact cellular morphology of white blood cells.
  • the first hemolytic agent is selected from the group consisting of quaternary ammonium salt cationic surfactants, alkyl ether ethoxy nonionic surfactants, alkanol polyoxyethylene ether surfactants, alkyl At least one of glycosides, triterpene saponins, steroidal saponins and diethylene glycol.
  • the first hemolytic agent includes diethylene glycol and formaldehyde dissolved in a buffer.
  • the antibody is CD45.
  • the present disclosure provides a method that utilizes the four-class leukocyte detection channel (i.e., DIFF channel) of a hematology analyzer to accurately distinguish primitive cells from leukocytes and perform accurate counting by only adding a fluorescently labeled antibody reagent.
  • DIFF channel the four-class leukocyte detection channel
  • a simple, low-cost method and blood cell analyzer that can detect primitive cells using a conventional blood cell analyzer.
  • a second object of the present disclosure is to provide a sample analyzer that draws a sample from a sample container through a sample distribution device and distributes at least part of the sample to a first reaction device and/or a second reaction device.
  • the sample, the antibody reagent, and the first hemolysis reagent are prepared to form a first sample to be tested; the sample and the first type of reagent are prepared through the second reaction device to form a second sample to be tested, and the first sample to be tested is transported through the first sample transport component
  • the test sample is transported from the first reaction device to the optical detection component for original cell detection, and the second test sample is transported from the second reaction device to the optical detection component through the first sample transport component for white blood cell classification and counting detection, thereby achieving
  • a sample analyzer can be used to detect the effects of blast cells and blood routine at the same time.
  • a third aspect of the present disclosure provides a sample analyzer, including a sample distribution device, an optical detection device, a first reaction device, a second reaction device, a first sample transport component and a controller;
  • the sample distribution device is used to draw a sample from the sample container and distribute at least part of the sample to the first reaction device and/or the second reaction device;
  • the first reaction device is at least used to provide a reaction site for a sample, an antibody reagent, and a first hemolytic reagent to prepare a first test sample, wherein the antibody in the antibody reagent can react with the surface antigen of the original cells in the sample.
  • the second reaction device is at least used to provide a reaction place for the sample and the first type of reagent to prepare and form the second sample to be tested;
  • the first sample transport component is used to transport the first sample to be tested from the first reaction device to the optical detection component, and is used to transport the second sample to be tested from the second sample to be tested.
  • Two reaction devices are delivered to the optical detection component;
  • the optical detection component includes a flow chamber, a light emitting component and a light receiving component.
  • the flow chamber is used for the first sample to be tested or the second sample to be tested to pass under the entrapment of the sheath liquid.
  • the light emitting component is used to emit light toward the first sample to be tested or the second sample to be tested in the flow chamber, and the light receiving component is used to receive the light emitted by the light emitting component through the third sample.
  • a first optical signal generated by a sample to be tested and used to receive the light a second optical signal generated by the light emitted by the emitting component passing through the sample to be tested;
  • the controller is configured to: analyze and obtain first particle information in the first test sample according to the first optical signal fed back by the light receiving component, where the first particle information includes original cell information. ;
  • the second particle information in the second test sample is analyzed and obtained.
  • the second particle information includes neutrophil information and eosinophil information. , monocyte information and lymphocyte information.
  • the sample analyzer further includes a housing, and the optical detection device, the first reaction device and the second reaction device are all located in the housing.
  • the controller is further configured to: during the sample distribution process of a single original cell detection item, control the sample volume distributed by the sample distribution device to the first reaction device to be less than or equal to 60 uL; and /Or, the controller is further configured to: during the sample distribution process of a single original cell detection item, control the sample amount distributed by the sample distribution device to the first reaction device to be between 50uL ⁇ 10uL.
  • the controller is further configured to: during the detection process of a single original cell detection item, control the reaction time between the sample and the antibody reagent in the first reaction device to be less than or equal to 90 seconds, preferably less than or equal to 90 seconds. Equal to 60s.
  • the antibody reagent is a CD45 reagent.
  • the sample distribution device is also used to absorb the antibody reagent from the first reagent container, and distribute the absorbed antibody reagent to the first reaction device; or, the sample analyzer It also includes a first reagent dispensing device, which is used to suck the antibody reagent from the first reagent container and distribute the sucked antibody reagent to the first reaction device.
  • the antibody reagent includes an antibody that is fluorescently labeled and capable of binding to an original cell surface antigen
  • the first optical signal includes irradiation of light emitted by the light emitting component to the first
  • the first side scattered light signal generated by the sample to be tested and the light emitted by the light emitting component are irradiated to the first sample to be tested and the first fluorescence signal generated by the fluorescent label; or, the antibody
  • the reagent contains an antibody that is not fluorescently labeled but can bind to the original cell surface antigen.
  • the second reaction pool is used to provide a reaction site for the sample, the antibody reagent, and the first fluorescent reagent to prepare and form the first reaction solution
  • the first optical signal includes a first side scattered light signal generated by the light emitted by the light emitting component irradiating the first test sample and the light emitted by the light emitting component irradiating the The first fluorescent signal generated by the first test sample via the first fluorescent reagent.
  • the first reaction device is used to provide a reaction place for the first reaction solution, the first hemolysis reagent, and the second fluorescent reagent to prepare and form the first test sample;
  • the first optical signal further includes a second fluorescent signal generated by the second fluorescent reagent when the light emitted by the light emitting component is irradiated to the first sample to be tested;
  • the controller analyzes and obtains the first particle information in the first sample to be tested based on the first optical signal fed back by the light receiving component: based on the first side scattered light signal and the third A fluorescence signal is analyzed to obtain the first original cell information and the first blood shadow area information, and the second blood shadow area information is obtained according to the second fluorescence signal analysis;
  • the first original cell information and the first blood shadow area information are subtracted from the second blood shadow area information to obtain the original cell information of the first test sample.
  • the second reaction device includes a third reaction pool and at least one of a fourth reaction pool and a fifth reaction pool.
  • the third reaction pool is used to combine the sample with the first reaction pool.
  • the reagent type provides a reaction place to prepare and form the second sample to be tested.
  • the fourth reaction pool is used to provide a reaction place for the sample and the second type reagent to prepare and form the third sample to be tested.
  • the fifth reaction pool The pool is used to provide a reaction place for the sample and the third type of reagent to prepare the fourth sample to be tested;
  • the first sample transport component is also used to transport the third sample to be tested or the fourth sample to be tested from the third sample to be tested.
  • Two reaction devices are delivered to the flow chamber;
  • the flow chamber is also used to allow the third sample to be tested or the fourth sample to be tested to pass under the entrapment of the sheath liquid, and the light emitting component is also used to face the third sample in the flow chamber.
  • the sample to be tested or the fourth sample to be tested emits light, and the light receiving component is also used to receive the third optical signal generated by the light emitted by the light emitting component through the third sample to be tested, or a fourth optical signal generated by the fourth sample to be tested;
  • the controller is further configured to perform at least one of the following parsing actions based on feedback information from the light receiving component:
  • the fourth particle information in the fourth sample to be tested is analyzed, and the fourth particle information includes reticulocyte information.
  • the controller when the test items of the sample include original cell test items and routine blood test items, the controller is configured to control the sample distribution device to perform the following actions in sequence: suck the sample from the sample container, Absorb the antibody reagent from the first reagent container, distribute the absorbed antibody reagent and part of the sample to the first reaction device, and distribute the absorbed part of the sample to the second reaction device, wherein the sample distribution device is from the The sample volume absorbed by the sample container at one time is greater than or equal to the sum of the sample volume distributed to the first reaction device and the sample volume distributed to the second reaction device; or,
  • the controller is configured to control the sample distribution device to perform the following actions in sequence: suck the sample from the sample container, and distribute part of the sample to The second reaction device absorbs the antibody reagent from the first reagent container, and distributes the absorbed antibody reagent and part of the sample to the first reaction device, wherein the sample distribution device absorbs the antibody reagent from the sample container in one go.
  • the sample amount is greater than or equal to the sum of the sample amount assigned to the first reaction device and the sample amount assigned to the second reaction device; or,
  • the controller is configured to control the sample distribution device to perform the following actions in sequence: draw the first sample from the sample container, draw the first sample from the first reagent Absorbing the antibody reagent from the container, distributing the absorbed antibody reagent and at least part of the first sample to the first reaction device, absorbing the second sample from the sample container, and distributing at least part of the absorbed second sample to the first reaction device second reaction device; or,
  • the controller is configured to control the sample distribution device to perform the following actions in sequence: suck the first sample from the sample container, and draw at least Distribute part of the first sample to the second reaction device, draw the second sample from the sample container, draw the antibody reagent from the first reagent container, and distribute the drawn antibody reagent and at least part of the second sample to the second reaction device.
  • First reaction device suck the first sample from the sample container, and draw at least Distribute part of the first sample to the second reaction device, draw the second sample from the sample container, draw the antibody reagent from the first reagent container, and distribute the drawn antibody reagent and at least part of the second sample to the second reaction device.
  • the sample dispensing device when the sample drawn by the sample dispensing device from the sample container is at least used for distributing to the first reaction device and the second reaction device respectively, the sample dispensing device takes the sample from the sample container.
  • the sample volume drawn by the above sample container at one time is less than or equal to 200uL.
  • the second reaction device at least includes a third reaction pool
  • the controller is further configured to: deduct the second particle from the original cell information of the first particle information. Abnormal lymphocyte data in the information to obtain measurement data of original cells; and/or,
  • the second reaction device at least includes a fourth reaction pool, and the controller is further configured to: deduct basophilicity in the third particle information from the original cell information of the first particle information. Granulocyte data to obtain the measurement data of original cells.
  • the controller is further configured to: during the sample distribution process of a single original cell detection item, control the amount of antibody reagent distributed by the sample distribution device to the first reaction device to be less than 5uL; and /or,
  • the controller is further configured to: during the dispensing process of the antibody reagent for a single original cell detection item, control the amount of the antibody reagent distributed to the first reaction device to be between 5uL ⁇ 2uL.
  • the sample analyzer further includes a hemoglobin detection device
  • the sample distribution device is also used to distribute at least part of the sample drawn to the hemoglobin detection device
  • the hemoglobin detection device is used to analyze at least the sample.
  • the hemoglobin parameters are measured with the fifth test sample made from the fourth type of reagent.
  • the controller is further configured to: analyze and obtain the hemoglobin parameters in the fifth test sample based on the information fed back by the hemoglobin detection device. Hemoglobin information; and/or,
  • the sample analyzer also includes an impedance detection device, the impedance detection device is used to measure a sixth test sample made of at least the sample and the fifth type of reagent, and the controller is also configured to: according to the The information fed back by the impedance detection device is analyzed to obtain platelet information and/or red blood cell information in the sixth sample to be tested.
  • the sample analyzer measures samples in a sample container and outputs at least the following two test results: original cell test results and routine blood test results; and/or,
  • the sample analyzer measures the sample drawn by the sample distribution device from the sample container at one time and outputs at least the following two test results: original cell test results and routine blood test results.
  • the sample analyzer further includes an identification device, the identification device is used to identify the sample container, and the controller is further configured to: according to the feedback information of the identification device, at least obtain the sample container
  • the test item information to be tested in the sample is controlled according to the test item information to be tested, and the sample analyzer is controlled to perform original cell testing and/or blood routine testing on the sample in the sample container; and/or,
  • the sample analyzer also includes a human-computer interaction device, which is configured with a primitive cell detection mode and a routine blood detection mode.
  • the controller is also configured to: select according to the operator's selection on the human-computer interaction device.
  • the detection mode controls the sample analyzer to perform original cell detection and/or routine blood detection on the sample in the sample container.
  • Figure 1 is a schematic diagram of the principle of using a flow cytometry blood cell analyzer to analyze white blood cells through nucleic acid fluorescence staining.
  • Figure 2 is a two-dimensional scatter plot of side light scattering intensity and nucleic acid fluorescence intensity obtained by detecting blood samples using the DIFF channel of a conventional blood cell analyzer.
  • Picture A is a two-dimensional scatter diagram of a normal (healthy) blood sample
  • B Pictured is a two-dimensional scatter plot of a blood sample containing primitive cells.
  • Figure 3 is a two-dimensional scatter plot of side light scattering intensity and fluorescence intensity obtained by using a blood cell analyzer to detect blood samples.
  • Picture A shows the side light scattering intensity-nucleic acid fluorescence intensity obtained by conventional white blood cell detection of normal blood samples.
  • Figure B is the side-light scattering intensity-antibody fluorescence intensity (SSC/antibody FL) obtained by using fluorescently labeled antibodies to detect blood samples containing original cells according to the disclosed method.
  • Three-dimensional scatter plot, in which the boxed area is the area where the original cells appear.
  • Figure 4 is a comparison of the two-dimensional scatter plot of side light scattering intensity and nucleic acid fluorescence intensity obtained by detecting blood samples using the DIFF channel of the blood cell analyzer of the present disclosure, and the two-dimensional scatter plot of side light scattering intensity and antibody fluorescence intensity.
  • Figure A shows the side-light scattering intensity of a sample containing primitive cells treated with a nucleic acid-containing fluorescent dye reagent under hemolytic conditions - the nucleic acid fluorescence intensity that should usually appear in the two-dimensional scatter plot of monocytes and lymphocytes.
  • Panel B shows a two-dimensional scatter plot of antibody fluorescence intensity-side light scattering intensity for the same sample treated with a fluorescently labeled CD45 reagent under hemolytic conditions. The boxed area is the location where primitive cells appear.
  • Figure 5 is a schematic flow chart of a primary cell detection method according to an embodiment of the present disclosure.
  • Figure 6 is a schematic flow chart of an original cell detection method according to another embodiment of the present disclosure.
  • Figure 7 is a schematic flow chart of an original cell detection method according to yet another embodiment of the present disclosure.
  • Figure 8 shows the third fluorescence intensity-side scattering intensity (FL/SSC) obtained by WNB channel detection. Schematic diagram of the distribution of each particle group in a dimensional scatter plot.
  • Figure 9 shows the two-dimensional scatter plot of antibody fluorescence intensity-lateral light scattering intensity (antibody/SSC) corresponding to the original cell particle population in the myeloid (Panel A) and lymphoid system (Panel B) samples respectively. Position in a two-dimensional scatter plot (panel C) of antibody fluorescence intensity versus forward light scatter intensity (antibody/FSC).
  • antibody/SSC antibody fluorescence intensity-lateral light scattering intensity
  • Figure 10 is a schematic diagram of the fluid circuit system of a blood cell analyzer provided by an embodiment of the present disclosure.
  • Figure 11 is a schematic diagram of the liquid path connection of the optical channel reaction cell, sampling component, optical detection unit, diluent supply device and fluid power device provided by this embodiment of the present disclosure.
  • Figure 12 is a schematic diagram of the fluid path connection between the impedance counting detection unit, the hemoglobin detection unit, the diluent supply device and the fluid power device provided by this embodiment of the present disclosure.
  • Figure 13 is a FL/SSC scatter plot of a sample containing a high content of immature granulocytes (IG) but no blast cells, detected according to Example 1.
  • Figure 14 is a FL/SSC scatter plot of a sample containing primitive cells obtained through detection according to Example 2.
  • Figure 15 is a WNB channel FL/SSC scatter plot (A) and a DIFF channel FL/SSC scatter plot (B) of a sample containing original cells detected according to Example 2.
  • Figure 16 is a CD45FL/SSC scatter plot of a sample containing primitive cells detected according to Example 4.
  • Figure 17 is a schematic diagram of the composition of a sample analyzer provided by another embodiment of the present disclosure.
  • Figure 18 is a schematic diagram of the working process of the sample needle provided by this embodiment of the present disclosure.
  • references to 'samples' in this article refer to blood samples, bone marrow samples or body fluid samples.
  • the blood sample can be peripheral blood or venous blood.
  • the body fluid sample is, for example, cerebrospinal fluid, pleural effusion, ascites fluid, myocardial fluid, synovial fluid, peritoneal dialysis fluid or fluid obtained by peritoneal washing.
  • Primitive cells as referred to herein, or “BLAST” as expressed herein, are used interchangeably and both refer to cells at the beginning stages of development of various blood cells. Primitive cells do not appear in the blood of healthy people, but people with some diseases, especially blood diseases, do.
  • hemolytic agent refers to an agent that can lyse red blood cells while allowing white blood cells to maintain their basic cellular shape and the cell membrane to be damaged to a certain extent.
  • hemolytic agents useful in the present disclosure may be selected from surfactants, alkyl glycosides, triterpene saponins, steroidal saponins, and the like.
  • Antibody refers to an antibody capable of binding to an antigen on the cell membrane surface of the original cell, unless otherwise specified. Antibodies, especially monoclonal antibodies. The antibody may bind non-specifically to the cell membrane surface antigens of the original cells, which means that the antibody can also bind to the cell membrane surface antigens of other cells in the blood, such as leukocytes. However, the binding conditions (such as firmness, quantity, location, etc.) of the antibodies to different blood cell surface antigens are different, and thus different signals can be obtained.
  • the 'fluorescently labeled antibody' mentioned herein refers to the above-mentioned antibody combined with a fluorescent label that can be used for detection.
  • the fluorescent markers can be selected from those fluorescent substances with appropriate excitation wavelengths according to different detection platforms.
  • the 'fluorescent dye' mentioned in this article refers to a dye that can specifically bind to intracellular substances (such as DNA, RNA or protein) and emit fluorescence of a specific wavelength under excitation by excitation light.
  • a fully automatic blood cell analyzer can be used to detect white blood cells in blood.
  • the leukocytes in the sample can be classified and counted through sheath flow impedance method, laser scattering method, flow cytometry technology combined with nucleic acid fluorescence staining, etc.
  • detection using flow cytometry technology (DIFF channel) combined with nucleic acid fluorescence staining uses nucleic acid fluorescence staining under hemolytic conditions and then analyzes through light signals.
  • the principle is shown in Figure 1. After the blood cell analyzer absorbs the blood sample, the sample is first treated with a hemolytic agent and a nucleic acid fluorescent dye. The red blood cells are destroyed and broken by the hemolytic agent, while the white blood cells still maintain their cell shape.
  • the nucleic acid fluorescent dye can enter the white blood cells and specifically bind to the nucleic acid in the cells. Then, using flow cytometry, the cells in the sample pass through the laser detection hole one by one. When the laser beam emitted by the light source irradiates the cells passing through the detection hole, scattered light is generated at various angles. At the same time, the fluorescent dye combined with the nucleic acid is emitted by the laser. Excited to emit fluorescence at a specific wavelength. According to the characteristics of the cells themselves (such as volume, staining degree, cell content size and content, cell nuclear density, etc.), the scattered light and fluorescence intensity generated at different angles are different. These scattered light and fluorescence signals can be detected after being received by the optical detector.
  • the forward light scattering intensity (Forward scatter, FS) reflects the volume of the cell
  • the side light scattering intensity (Side scatter, SS) reflects the complexity of the internal structure of the cell
  • the fluorescence intensity (Fluorescence, FL) reflects the nucleic acid substances in the cell (including DNA and RNA) content.
  • These parameters can be used to classify and count white blood cells in a sample.
  • the two-dimensional signal data of side light scattering intensity and fluorescence intensity are used to form a two-dimensional scatter plot in the DIFF channel, and four-category counts of lymphocytes, monocytes, eosinophils and neutrophils can be obtained (see figure Figure A in 2).
  • blast-sensitive cells When there are blast cells in the patient's peripheral blood, the blast cells are also stained with fluorescent dyes, and their particle information often covers the lymphocyte and monocyte areas in the SS-FL two-dimensional scatter plot (called "blast-sensitive cells"). area", see panel B in Figure 2), resulting in the inability to distinguish lymphocytes, monocytes, and primitive cells, and thus lymphocytes and monocytes cannot be accurately quantified.
  • current commercial blood cell analyzers can only provide alarm prompts for primitive cells but cannot distinguish and accurately count primitive cells.
  • the sensitivity and specificity of alarm prompts are not high, and false positives and false negatives may exist.
  • FIG. 4 a comparison is made between the two-dimensional scatter plot obtained by the conventional nucleic acid fluorescence method for leukocyte analysis and the two-dimensional scatter plot obtained by analyzing the method of the present disclosure when primitive cells are present in the sample.
  • Picture B shows a two-dimensional scatter plot of antibody fluorescence intensity-side light scattering intensity after treating a sample containing primitive cells with a fluorescently labeled antibody reagent under hemolytic conditions. The boxed area is the location where primitive cells appear. .
  • Picture A shows the area where monocytes and lymphocytes should usually be distinguished after processing the same sample with nucleic acid-containing fluorescent dye reagents under hemolytic conditions. It is completely impossible to distinguish between blasts, monocytes and lymphocytes.
  • the inventor proposed a blood cell analyzer and a blast cell detection method that can distinguish blast cells from other white blood cells and accurately count blast cells.
  • a sample analysis method is provided.
  • the method includes receiving operating instructions and performing original cell detection on the sample.
  • the operation instruction can be an instruction selected by the user (for example, for a sample that is known to contain primitive cells, you can directly choose to perform primitive cell detection; or after the instrument issues a primitive cell alarm, you can also choose whether to perform primitive cell detection), or it can be a blood cell It is directly triggered by the original cell alarm issued by the analyzer during detection.
  • Performing original cell detection on the sample may include the following steps.
  • absorbing a sample includes absorbing at least part of the sample as a first sub-sample.
  • S20 process the sample, including mixing the first sub-sample with a first type of reagent to obtain a first test sample; wherein the reagent includes a first reagent and a second reagent, and the first reagent includes a first hemolysis reagent, the mixing time of the first hemolysis reagent and the first subsample does not exceed 2 minutes, and the second reagent includes a fluorescently labeled antibody, and the antibody can bind to the surface antigen of the original cell.
  • Detecting the sample to be tested includes causing the particles in the first sample to be tested to pass through the detection area of the optical detection device one by one, and using the light source of the optical detection device to detect the particles in the first sample to be tested.
  • the particles are irradiated to measure the light scattering signal and the fluorescence signal of the first test sample.
  • the light scattering signal includes the first side light scattering intensity signal from the first test sample.
  • the fluorescence signal includes the first side light scattering intensity signal from the first test sample.
  • the antibody fluorescence intensity signal of the fluorescently labeled antibody is causing the particles in the first sample to be tested to pass through the detection area of the optical detection device one by one, and using the light source of the optical detection device to detect the particles in the first sample to be tested.
  • the particles are irradiated to measure the light scattering signal and the fluorescence signal of the first test sample.
  • the light scattering signal includes the first side light scattering intensity signal from the first test sample.
  • the fluorescence signal
  • obtaining particle information including obtaining particle information in the first subsample according to the first side light scattering intensity signal and the antibody fluorescence intensity signal, where the particle information in the first subsample includes original cells information.
  • This method can utilize the DIFF channel of the hematology analyzer detailed below, or it can add new separate detection channels.
  • a part of the sample in the step of aspirating the sample in S10, can be quantitatively aspirated through the sample aspirating device as the first sub-sample for subsequent primary cell detection.
  • the step of aspirating the sample may also include aspirating part of the sample multiple times to obtain multiple subsamples for different tests, such as leukocyte analysis, platelet analysis, hemoglobin analysis, and so on. This step may also include dividing the sample drawn once into several sub-samples for different detections.
  • the first subsample is mixed with the first and second reagents to perform hemolysis and antibody-antigen reaction to prepare the first test sample.
  • the first and second reagents may be mixed with the sample one after the other in any order, or may be mixed with the sample simultaneously.
  • the mixed sample is incubated for a certain period of time to break up the red blood cells and allow fluorescently labeled antibodies to bind to antigens on the cell surface.
  • the first reagent containing the first hemolytic agent in this step can quickly hemolyze red blood cells. For example, hemolysis can be completed in about 2 minutes. Usually the hemolysis time is within 1 minute, more usually 30 to 80 seconds, or even 50 to 60 seconds. .
  • the reaction between the fluorescently labeled antibody in the second reagent and the antigen on the cell surface usually takes about 45 to 60 seconds at a temperature of about 42 to 45°C. The reaction time will vary depending on the antibody selected and the incubation temperature.
  • the first hemolytic agent in the first reagent may be the same hemolytic agent used in the DIFF channel in conventional leukocyte detection, or may be different from the hemolytic agent used in the DIFF channel.
  • the first hemolytic agent may be one hemolytic agent or a combination of multiple hemolytic agents.
  • the first hemolytic agent may include cationic surfactants selected from quaternary ammonium salts, alkyl ether ethoxy non-ionic surfactants, alkanol polyoxyethylene ether surfactants, alkyl glycosides, triglycerides, etc.
  • At least one specific example of terpene saponins, steroid saponins and diethylene glycol may be a glycoside compound of general formula I: R-(CH 2 ) n -CH 3 (I)
  • R is selected from the group consisting of monosaccharides, deoxymonosaccharides and polysaccharides, and n is an integer from 5 to 17.
  • the hemolytic agents disclosed in Chinese patent application CN111602052A can be used as the first hemolytic agent of the present disclosure. The entire contents of this patent document are incorporated herein by reference.
  • the first hemolytic agent may include diethylene glycol, preferably further including formaldehyde which has a fixation effect on cells.
  • the first hemolytic agent may include a cationic surfactant, a nonionic surfactant, and and aromatic organic acids and/or their salts.
  • the aromatic organic acid can be selected from the group consisting of terephthalic acid, phthalic acid, hydroxybenzoic acid, acetylsalicylic acid, p-aminobenzoic acid, benzenesulfonic acid, p-toluenesulfonic acid and hydroxybenzenesulfonic acid. one or more of them.
  • the cationic surfactant is selected from at least one of quaternary ammonium salts and pyridinium salts.
  • the nonionic surfactant is selected from at least one type of polyoxyethylene surfactant, Spann and Tween.
  • the first reagent may also contain buffers (such as phosphoric acid and its salts, citric acid and its salts, acetic acid and its salts, etc.), preservatives (such as sodium azide, ProClin series, etc.), metal chelating agents (such as EDTA sodium salt), osmotic pressure regulator (such as sodium chloride, etc.) and other components.
  • buffers such as phosphoric acid and its salts, citric acid and its salts, acetic acid and its salts, etc.
  • preservatives such as sodium azide, ProClin series, etc.
  • metal chelating agents such as EDTA sodium salt
  • osmotic pressure regulator such as sodium chloride, etc.
  • the available first reagent may be a commercialized hemolytic reagent for leukocyte detection, such as M-68P LD hemolytic reagent.
  • Other hemolytic agents may also be used, such as diethylene glycol and formaldehyde dissolved in a buffer solution.
  • the fluorescently labeled antibodies in the second reagent may be those capable of binding to the original cell surface antigen and linked to a suitable fluorescent label, especially a monoclonal antibody.
  • the present disclosure has no special restrictions on antibodies. As long as they have different binding abilities to various types of cells and can distinguish primitive cells from other white blood cells through antibody fluorescence and side light scattering intensity, they can be used. For example, they can be one of CD45, CD71, and CD34. species, or a combination of two or more species. Preferably only one antibody is used, more preferably the antibody is CD45.
  • the disclosure does not have any special restrictions on the fluorescent label, as long as it does not affect the binding of the antibody to the antigen after binding to the antibody, nor does it affect the use of other reagents (such as hemolytic reagents).
  • Those fluorescent substances that can be excited by the light source can be selected as markers according to the wavelength of the laser light source used by the blood cell analyzer. These fluorescent markers are well known to those skilled in the art. Examples of fluorescent markers include FITC (blue light excitation) and APC (red light excitation), but are not limited thereto.
  • Commercially available fluorescently labeled antibodies can be used, such as CD45-FITC, CD45-APC, etc.
  • the concentration of the fluorescently labeled antibody in the second reagent can be 100-200 ⁇ g/mL, and the solvent is usually a phosphate buffer with a pH of 7.2.
  • the second reagent may also include sodium azide and bovine serum albumin.
  • the volume mixing ratio of the first reagent to the sample can be the volume used in conventional hemolysis processing, such as 50:1. For example, approximately 1 mL of the first reagent may be mixed with approximately 20 ⁇ L of sample. The volume mixing ratio of the second reagent to the sample may be 1:20.
  • the first test sample that has completed red blood cell hemolysis and antibody-antigen reaction is transported to the optical detection unit for optical detection.
  • each particle passes through the detection area of the flow chamber one by one. Under the irradiation of the laser light source, scattered light in different directions is generated.
  • the fluorescent markers carried by the antibodies are excited and emit fluorescence.
  • the light scattering signal and fluorescence signal of each particle are collected by signal collection devices arranged around the detection zone and transmitted to the processor.
  • the light scattering signal of the first test sample at least includes a first lateral light scattering intensity signal.
  • the first forward light scattering intensity signal can also be collected at the same time, and even light scattering intensity signals at other angles can be further collected as needed.
  • the fluorescence signal of the first test sample is the antibody fluorescence intensity signal emitted by the fluorescent label carried by the antibody.
  • the detection of primitive cells can utilize the DIFF channel of conventional leukocyte detection, and only the fluorescent staining reagent used for leukocyte detection is replaced by an antibody fluorescent reagent.
  • the detection of primitive cells can be performed using a separate antibody channel, while the leukocytes are still detected using the DIFF channel.
  • a two-dimensional scatter plot of SSC-antibody FL can be formed according to the received first side light scattering intensity signal and fluorescence intensity signal, and particles of the first subsample including original cell information are obtained. information. Still referring to Figure 3 and Figure 4 B, it can be seen that in the coordinates containing the antibody fluorescence signal, the particle clusters of original cells can be significantly distinguished from other white blood cell particle clusters, so that accurate original cell information can be obtained. For samples from healthy subjects, no particles may be detected in the area of the original cell particle cluster (i.e., the percentage of original cells is 0%) or the number of particles may be extremely low. For samples from patients (shown in Figure 4, panel B), accurate primary cell classification and counting can be further obtained.
  • the method can also eliminate the influence of primitive cells on the detection of white blood cells and further obtain an accurate count of white blood cells, which can be implemented in a variety of embodiments.
  • the DIFF channel of the hematology analyzer can be used for one-time sample preparation and one-time detection to obtain information on primitive cells and white blood cells at the same time, and can obtain accurate primitive cell counts, and obtain four-differentiation counts of white blood cells without being affected by the sample. effects on primitive cells.
  • This embodiment is particularly suitable for blood cell analyzers equipped with dual fluorescence detectors.
  • the fluorescence wavelength emitted by the fluorescent label labeling the antibody should be different from the fluorescence wavelength emitted by the fluorescent reagent used in leukocyte counting.
  • the method for detecting primary cells may include the following steps.
  • step S110 Take part of the sample as the first sub-sample. This step is the same as step S10 in the above embodiment.
  • step S120 process the sample, mix the first subsample with a first reagent including a first hemolytic reagent, a second reagent including a fluorescently labeled antibody, and a third reagent including a first fluorescent dye to obtain a first test sample.
  • a first reagent including a first hemolytic reagent a second reagent including a fluorescently labeled antibody
  • a third reagent including a first fluorescent dye to obtain a first test sample.
  • the first sub-sample is also mixed with a third reagent including a first fluorescent dye.
  • the first, second and third reagents can be mixed with the first subsample sequentially in any order or simultaneously.
  • the first and second reagents are as described above, and the mixing method with the sample is also as described above.
  • the third reagent may be a nucleic acid fluorescent dye reagent commonly used for DIFF channel leukocyte detection.
  • the first fluorescent dye in the third reagent is a nucleic acid fluorescent dye.
  • Commercial nucleic acid fluorescent dyes and nucleic acid-specific fluorescent dyes that have been disclosed in some patent applications can be used in the method.
  • Thermofisher's SYTO series of nucleic acid dyes can be cited.
  • the fluorescent dyes disclosed in Chinese Patent Application No.: CN201010022414.6, the anthocyanin dyes disclosed in Chinese Patent Application No.: CN200910109215.6, the fluorescent dyes disclosed in CN200810216864.1, etc. can all be used in this disclosure.
  • the entire contents of the above patent documents are incorporated herein by reference.
  • the first fluorescent dye can be dissolved in a suitable organic solvent (such as ethanol, methanol, ethylene glycol, etc.) to form the third reagent.
  • a suitable organic solvent such as ethanol, methanol, ethylene glycol, etc.
  • the third reagent can be those that are commercially available, such as M-68P FD staining solution.
  • the present disclosure has no particular limitations on the third reagent.
  • the volume mixing ratio of the third reagent and the sample can be the mixing ratio used in conventional white blood cell detection using the DIFF channel, for example, it can be 1:1.
  • the red blood cells have been broken, and the blood cells that still maintain intact cell shapes, such as white blood cells, and possible primitive cells, have been stained for intracellular nucleic acid substances, and fluorescent markers have also been combined with the cell membrane. of antibodies.
  • the sample to be tested detects the sample to be tested, which is the same as the above embodiment.
  • the particles in the first sample to be tested pass through the detection area of the flow chamber one by one. Under the irradiation of the laser light source, scattered light in different directions is generated. At the same time, due to the cell surface binding When the antibody is added, the fluorescent label carried by the antibody is excited and emits antibody fluorescence. The difference is that the nucleic acid inside the cell is combined with the first fluorescent dye. This dye has the characteristic of significantly increasing the fluorescence quantum yield after binding to the nucleic acid, so it is also known as Excited to emit fluorescence.
  • the light scattering signal of the first test sample at least includes a first lateral light scattering intensity signal.
  • the first forward light scattering intensity signal can also be collected at the same time, and even light scattering intensity signals at other angles can be further collected as needed.
  • the fluorescence signal of the first test sample is the antibody fluorescence intensity signal emitted by the fluorescent label carried by the antibody and the first fluorescence intensity signal emitted by the first fluorescent dye bound to the nucleic acid.
  • S140 Obtain particle information, including original cell information obtained based on the first side light scattering intensity signal and the antibody fluorescence intensity signal; and obtain white blood cell information based on the first side light scattering intensity signal and the first fluorescence intensity signal.
  • the cells simultaneously bind the fluorescently labeled antibody and the first fluorescent dye, signals in at least four dimensions can be measured for each particle, and signals in different dimensions can be used to obtain information about more particles.
  • accurate classification and counting of original cells can be directly obtained using the first side-light scattering intensity signal and the antibody fluorescence intensity signal.
  • the white blood cells obtained using the first side light scattering intensity signal and the first fluorescence intensity signal The information is affected by the possible presence of primitive cells in the sample, making it difficult to distinguish between lymphocytes, monocytes, and primitive cells (see Figure 4, Panel A).
  • the influence of the original cells is deducted from the particle information of the first sub-sample (that is, using the first side light scattering After the intensity signal and antibody fluorescence intensity signal divide the information of the original cell particles, the original cell information is deducted from the scatter plot of the first side light scattering intensity signal and the first fluorescence intensity signal) to obtain neutrophils, eosinophils Differential counting of granulocytes, monocytes and lymphocytes (S142).
  • steps S141 and S142 immature granulocyte information and/or atypical lymphocyte information can be further obtained, if these cells exist in the sample.
  • test samples are prepared respectively, one for blast cell detection, one for four-classification leukocyte detection in DIFF channel, and one for nucleated red blood cells and basophils in WNB channel. Detection of cells and lymphocytes.
  • This embodiment is particularly suitable for blood cell analyzers equipped with only one fluorescence detector.
  • the method for detecting primary cells may include the following steps.
  • sample draw the sample as the first, second and third sub-sample.
  • the sample can be aspirated once and divided into three blood segments for separate detection; the sample can also be aspirated several times.
  • S220 process each subsample separately. In this step, the three subsamples are processed separately.
  • the first sub-sample is mixed with the above-mentioned first reagent and second reagent to obtain the first test sample.
  • This step is the same as step S20 in the above embodiment.
  • the fourth reagent includes a second hemolytic reagent
  • the fifth reagent includes a second fluorescent dye
  • the sixth reagent includes a third hemolytic reagent.
  • the second hemolytic agent may be the same as or different from the first hemolytic agent
  • the third hemolytic agent may be different from both the first and second hemolytic agents.
  • the second fluorescent dye is different from the first fluorescent dye.
  • the fifth and sixth reagents are fluorescent reagents and hemolytic reagents commonly used for WNB channel detection.
  • the fourth reagent includes a second hemolytic agent.
  • the second hemolytic reagents are those used in the DIFF channel in conventional white blood cell detection.
  • the first hemolytic agent may be one hemolytic agent or a combination of multiple hemolytic agents.
  • the first hemolyzing agent can be a surfactant, such as a cationic surfactant (such as a quaternary ammonium salt cationic surfactant), a nonionic surfactant (such as an alkanol polyoxyethylene ether surfactant) .
  • the first hemolytic agent may be an alkyl glycoside, a triterpene saponin, or a steroidal saponin. Specific examples may be glycoside compounds of general formula I: R-(CH 2 ) n -CH 3 (I)
  • R is selected from the group consisting of monosaccharides, deoxymonosaccharides and polysaccharides, and n is an integer from 5 to 17.
  • the hemolytic agents disclosed in Chinese patent application CN111602052A can be used as the first hemolytic agent of the present disclosure. The entire contents of this patent document are incorporated herein by reference.
  • the fourth reagent may also contain components such as buffers, preservatives, metal chelating agents, and osmotic pressure regulators.
  • the available first reagent may be a commercialized hemolytic reagent for leukocyte detection, such as M-68P LD hemolytic reagent.
  • the volume mixing ratio of the fourth reagent and the sample is the ratio used in conventional DIFF channel detection, for example, it can be 50:1.
  • the fifth reagent includes a second fluorescent dye.
  • the second fluorescent dye is a type of dye that can bind to proteins. Examples of the second fluorescent dye may be compounds with the following formula:
  • R 7 , R 8 and R 9 can be selected from H, halogen, cyano, hydroxyl, C 1-18 alkyl, C 1-18 alkylsulfonate, sulfonate and C 1-15 alkyl COOR 10 , Wherein R 10 is H or C 1-6 alkyl;
  • R 5 and R 6 are each independently selected from C 1-18 alkyl COOR 11 , C 1-18 alkyl OR 11 and benzyl group, wherein the benzyl group can be selected from halogen, hydroxyl, mercapto, cyano, nitro, alkyl Substituted with groups from aryl, aryl, alkoxy, heterocyclyl, haloalkyl, amino, alkylamino, amido and carboxyl groups, and R 5 and R 6 are not benzyl groups at the same time;
  • Each occurrence of R 11 can be independently H, C 1-18 alkyl or phenyl, where phenyl can be selected from halogen, hydroxyl, mercapto, cyano, nitro, alkyl, aryl, alkoxy , substituted by heterocyclyl, haloalkyl, amino, alkylamino, amide and carboxyl groups;
  • X is -CH 2 -, -C(CH 3 ) 2 -, -O-, -S- or -Se-;
  • Z - is an anion
  • the second fluorescent dye can be dissolved in a suitable solvent (such as methanol, ethanol, ethylene glycol, etc.) at a concentration of, for example, 50 ⁇ g/mL as the fifth reagent.
  • a suitable solvent such as methanol, ethanol, ethylene glycol, etc.
  • dyes used as second fluorescent dyes may be those already commercialized, such as M-68P FN staining solution.
  • the sixth reagent includes a third hemolytic agent.
  • the third hemolytic agent is different from the first and second hemolytic agents.
  • the third hemolytic agent may include at least one surfactant selected from cationic surfactants and nonionic surfactants.
  • the cationic surfactant can be selected from quaternary ammonium salt type cationic surfactants having the structure shown in Formula 1.
  • the structural formula of specific embodiments is:
  • R 1 is an alkyl or alkenyl group with 6 to 14 C atoms
  • R 2 and R 3 are alkyl or alkenyl groups with 1 to 4 C atoms;
  • R 4 is an alkyl group, alkenyl group or benzyl group with 1 to 4 C atoms
  • B is a halogen atom.
  • the above-mentioned cationic surfactant is preferably dodecyltrimethylammonium chloride, cetyltrimethylquaternary ammonium bromide, or stearyldimethylbenzylquaternary ammonium chloride.
  • the nonionic surfactant can be selected from the group consisting of octylphenyl polyoxyethylene ether and polyoxyethylene cetyl ether.
  • the third hemolytic agent may also include aromatic organic acids or salts thereof, such as salicylic acid and its salts, benzoic acid and its salts.
  • the third hemolytic agent as the sixth reagent can be a commercial hemolytic agent used for leukocyte detection, such as (M-68P LN hemolytic agent).
  • the third hemolytic agent can be an acidic hemolytic agent compared with the first or second hemolytic agent. agent (pH3.0).
  • the sixth reagent is similar to the first reagent and may also include a buffer, a preservative, an osmotic pressure regulator, a complexing agent, etc.
  • the volumetric mixing ratio of the fifth and sixth reagents to the sample can be in accordance with the way a conventional WNB channel processes samples.
  • the mixing ratio of the fifth reagent and the sample can be 1:1
  • the mixing ratio of the sixth reagent and the sample can be 50:1.
  • the subsamples can be processed sequentially in the same mixing chamber or in different mixing chambers.
  • S230 detect each sample to be tested separately. In this step, three samples to be tested are tested separately.
  • S231 detects the first sample to be tested. This step is the same as step S30 in the previous embodiment.
  • this detection step three samples to be tested are detected by the optical detection unit in sequence, thereby obtaining optical signals of particles in each sample to be tested. These optical signals are transmitted to the processor for the next step.
  • the original cell information is obtained based on the first side light scattering intensity signal and the antibody fluorescence intensity signal, which is the same as step S40 in the previous embodiment.
  • S243 For the third test sample, obtain particle information including at least lymphocytes, basophils and nucleated red blood cells based on the third side light scattering intensity signal and the third fluorescence intensity signal. This step is the same as the detection step in conventional sample analysis using the WNB channel.
  • differential counts of basophils and nucleated red blood cells can be obtained, in addition to accurate lymphocyte counts. Due to the characteristics of the second fluorescent dye used, even if there are primitive cells in the sample, they will not interfere with the differential counting of lymphocytes.
  • Figure 8 which shows a schematic diagram of the distribution of each particle group in the two-dimensional scatter plot of fluorescence intensity-lateral light scattering intensity obtained by WNB channel detection. As can be seen from Figure 8, the location of primitive cells overlaps with abnormal lymphocytes, but can be well distinguished from normal lymphocytes. Therefore, an accurate lymphocyte percentage for the third test sample can be obtained in the WNB channel. In addition, the particle populations of basophils and nucleated red blood cells are not affected by primitive cells, so accurate differential counts of basophils and nucleated red blood cells can be obtained.
  • the second test sample obtain particle information including at least lymphocytes, monocytes, neutrophils and eosinophils based on the second side light scattering intensity signal and the second fluorescence intensity signal.
  • the classification of lymphocytes and monocytes among the four categories of white blood cells may be interfered by primitive cells, making it impossible to perform classification and counting.
  • the primitive cell information obtained from the first test sample and the lymphocyte information obtained from the third test sample are combined to eliminate
  • the influence of the original cell information on the particle information in the second subsample is used to obtain classified counts of neutrophils, eosinophils, monocytes and lymphocytes.
  • steps S2421 and S2422 immature granulocyte information and/or atypical lymphocyte information can be further obtained, if these cells exist in the sample.
  • the blast cells in the sample are myeloid blast cells or lymphoid blast cells.
  • Figure 9 shows the two-dimensional scatter plot of antibody fluorescence intensity-lateral light scattering intensity (antibody/SSC) from the original cell particle population in myeloid and lymphoid samples respectively corresponding to the antibody fluorescence intensity-front Position in a two-dimensional scatter plot of light scattering intensity (antibody/FSC).
  • the primitive cells can be further determined to be myeloid and/or lymphoid primitive cells based on the antibody fluorescence intensity signal and the forward light scattering intensity signal.
  • the first forward light scattering block signal and the fluorescence intensity signal of the particles of the first test sample can be further used to determine whether the primitive cells in the tested sample are myeloid and/or lymphoid primitive cells. ; or alarm for the presence of lymphoid blasts and/or myeloid blasts in the sample being tested.
  • the primitive cells in the sample to be tested can be directly counted without determining whether there are primitive cells.
  • an alarm is issued that primitive cells are present in the sample to be tested, or a primitive cell count value is output.
  • lymphocytes including lymphocytes can be well distinguished.
  • Cell classification information for cells, monocytes, neutrophils, and eosinophils see Figure 3, Panel B).
  • the original cell detection method of the present disclosure also includes obtaining the leukocyte classification information in the sample based on the first side light scattering intensity signal and the antibody fluorescence intensity signal obtained from the first test sample, so
  • the white blood cell information includes information about lymphocytes, monocytes, neutrophils and eosinophils.
  • the method includes differential counting of lymphocytes, monocytes, neutrophils and eosinophils.
  • the sample analysis method further includes performing routine testing on the sample.
  • the conventional testing includes analyzing and classifying and counting various cells in the sample to be tested, and also includes analyzing some biochemical indicators.
  • the routine test may be a routine blood test in the clinic.
  • the routine detection may include detection and analysis of red blood cells, white blood cells, platelets, hemoglobin and other items.
  • routine test finds that primitive cells may be present in the sample, you can choose to initiate the detection of primitive cells; if the routine test does not find that primitive cells may be present in the sample, you do not need to initiate the detection of primitive cells.
  • This embodiment is suitable for subjects who are not confirmed to be sick or are not suspected to be sick, such as a large number of general outpatients or patients undergoing health examinations, so as to save the time and cost of testing.
  • the sample analysis method of the present disclosure includes performing leukocyte detection using a DIFF channel on the sample.
  • the routine detection includes the following steps.
  • At least part of the sample is taken as a first initial subsample.
  • This step includes mixing the first initial sub-sample with a third reagent and a fourth reagent to obtain a first initial test sample.
  • the third reagent and fourth reagent are as described above.
  • the particles in the first initial test sample are made to pass through the detection area of the optical detection unit one by one and the light source of the optical detection unit is used to irradiate the particles in the first initial test sample to obtain the first initial test sample.
  • the light scattering signal and fluorescence signal of the particles in the test sample includes the first initial side light scattering intensity signal from the first initial test sample; the fluorescence signal includes the first fluorescent dye from the first initial fluorescence signal.
  • the particle information of the first initial test sample according to the first initial side light scattering intensity signal and the first initial fluorescence intensity signal where the particle information at least includes white blood cell information; and when the white blood cell information is abnormal , issuing a primitive cell alarm.
  • the particle populations of lymphocytes and monocytes overlap with the primitive cell particle population in the two-dimensional scatter plot of fluorescence intensity-side light scatter intensity. , resulting in the inability to classify lymphocytes and monocytes.
  • an alarm will be issued in response to this situation that there may be primitive cells in the sample.
  • the user can choose to issue an operation instruction, or the alarm directly triggers the operation instruction, and the original cells are detected according to any one of the methods in the above embodiments.
  • the sample analysis method of the present disclosure includes performing four-classification of leukocyte detection using the DIFF channel and nucleated red blood cells, basophils and lymphocytes using the WNB channel on the sample.
  • the routine detection includes the following steps.
  • At least part of the sample is taken as a first initial subsample and a second initial subsample respectively.
  • processing the first and second initial sub-samples respectively including mixing the first initial sub-sample with the third reagent and the fourth reagent to obtain the first initial test sample, and mixing the second initial sub-sample with the fifth reagent.
  • the third, fourth, fifth and sixth reagents are as described above respectively.
  • the particles in the first and second initial samples to be tested are passed through the detection area of the optical detection unit one by one, and the light source of the optical detection unit is used to detect the particles in the first and second initial samples to be tested respectively.
  • the particles are irradiated to obtain light scattering signals and fluorescence signals of the particles in the first and second initial test samples, and the light scattering signals include the first initial lateral light from the first initial test sample.
  • the fluorescence signal includes the first initial fluorescence signal from the first fluorescent dye in the first initial test sample and the first initial fluorescence signal from the first test sample.
  • the particle information of the first initial test sample is obtained according to the first initial side light scattering intensity signal and the first initial fluorescence intensity signal, where the particle information at least includes initial white blood cell information, and according to the first Two initial side-light scattering intensity signals and a second initial fluorescence intensity signal obtain particle information of the second initial test sample, the particle information at least includes initial lymph information; and when the initial white blood cell information is abnormal, send out Primitive cell alarm.
  • the primary cell detection of any of the above embodiments can be performed.
  • the original cell detection as in the embodiment shown in FIG. 5 can be performed.
  • the primitive cell information obtained in the primitive cell detection and the initial lymphocyte information obtained in the routine detection are combined to eliminate the influence of the primitive cells on the initial white blood cell information obtained in the routine detection and obtain neutrophils. Differential counts of cells, eosinophils, monocytes, and lymphocytes. Furthermore, information on immature granulocytes and/or atypical lymphocytes can be obtained if present in the sample.
  • the present disclosure further provides a blood cell analyzer.
  • a blood analyzer including a sample suction device, a reagent supply device, a mixing chamber, a light source, an optical flow chamber, an optical detector, a processor, and a non-transitory computer-readable storage medium programmed with a computer application.
  • the sample suction device is used to suck the sample and transport the sucked sample to the mixing chamber.
  • a part of the sample may be drawn as needed to be used for one or several tests in each embodiment of the above sample analysis method.
  • the sample suction device can suck a certain amount of samples at one time and divide them into several sub-samples for sample processing and testing respectively; it can also suck a certain amount of samples multiple times and use them as sub-samples for separate testing. Processing and testing.
  • the reagent supply device is used to supply reagents and deliver them to the mixing chamber. There may be multiple reagent supply devices, each of which is used to provide different reagents. As described in the above embodiments of the sample analysis method, the reagent supply device is used to provide, for example, one or more of the first to sixth reagents required for sample processing according to different detection needs.
  • the mixing chamber is used for mixing and reaction of samples and reagents. There can also be multiple mixing chambers to process each subsample required for different tests. In addition, the same mixing chamber can also process different subsamples one after another.
  • the mixing chamber can be heated to the appropriate reaction temperature for optimal sample processing.
  • the sample to be tested is prepared after the sample is processed with corresponding reagents in the mixing chamber.
  • the sample to be tested is transported to the detection unit through the pipeline assembly.
  • the sample to be measured is transported to the optical detection unit.
  • the light source, optical flow chamber, detection hole and optical detector are all components of the optical detection unit.
  • the light source is used to aim the light beam at the detection hole of the optical flow chamber connected to the mixing chamber.
  • the light source is usually a laser, and its wavelength can be in the blue or red range.
  • the fluorescent label in the fluorescently labeled antibody suitable for different wavelength light sources can be selected to match the reagents configured in the blood cell analyzer used (such as the third reagent containing the first fluorescent dye and/or the third reagent containing the second fluorescent dye).
  • the fifth reagent of dye performs the above sample analysis.
  • the optical detector is used to detect the light scattering signal and fluorescence signal of the particles in the sample to be tested that pass through the detection hole one by one.
  • the light scattering signal includes the forward light scattering intensity signal collected by the forward scattered light signal collection device disposed on the optical axis, and the lateral light collected by the side scattered light signal collection device disposed on the side of the optical axis. Scatter intensity signal.
  • the optical detector also includes a fluorescence signal collection device arranged on the side of the optical axis to collect fluorescence intensity signals.
  • the fluorescence signal collection device can collect the antibody fluorescence intensity signal emitted from the fluorescently labeled antibody that has been bound to the cell membrane surface, and/or collect the fluorescence signal from the fluorescent dye that enters the interior of the cell and binds to nucleic acid or protein (for example, The fluorescence intensity signal emitted by the first or second fluorescent dye).
  • the light scattering signal and fluorescence signal of the particles detected by the optical detector are transmitted to the processor for analysis.
  • the processor is operatively connected to the optical detector, and when the computer application program in the non-transitory computer-readable storage medium is executed by the processor, the mixing of the reagent and the sub-sample in the mixing chamber is controlled. time, and the processor obtains corresponding particle information based on the received light scattering signal and volume fluorescence signal, and executes it for each detection method (such as any original cell detection method and/or conventional detection method in the above embodiments) Steps to obtain particle information.
  • the steps for obtaining particle information are the same as in the previous methods and will not be described again here.
  • the cell analyzer may further include a hemoglobin detection unit and an impedance counting detection unit.
  • the hemoglobin detection unit can be used for hemoglobin detection.
  • the impedance counting detection unit can be used for detection of red blood cells and/or platelets.
  • FIGS. 10 to 12 The following describes a blood cell analyzer provided by specific embodiments of the present disclosure with reference to FIGS. 10 to 12 . It should be understood by those skilled in the art that the blood cell analyzer used in the present disclosure is not limited to the embodiments described in detail below.
  • the blood cell analyzer includes a sample suction device 100, an optical channel mixing chamber 200, an optical detection unit 300, a hemoglobin detection unit 400, an impedance counting detection unit 500, a first reagent supply device 600, a second reagent supply device 700, Transfer pipeline assembly 800, fluid power device 900, waste liquid tank 1000, diluent supply device 1100, and a processor and a non-transitory computer-readable storage medium (not shown) programmed with a computer application program, aspiration device 100 , optical channel mixing chamber 200, optical detection unit 300, hemoglobin detection unit 400, impedance meter
  • the detection unit 500, the first reagent supply device 600, the second reagent supply device 700, the waste liquid pool 1000, the diluent supply device 1100 and the fluid power device 900 are connected through the transport pipeline assembly 800 to form a liquid circuit system.
  • the fluid power device 900 includes a first syringe 910 and a second syringe 920 , and the measurement range of the first syringe 900 is smaller than the measurement range of the second syringe 920 .
  • the first syringe 910 is multiplexed to provide power for sampling, blood separation (sampling), and sample ejection (including optical channel sample ejection and sheath flow impedance sample ejection).
  • the second syringe 920 is reused to provide power for sample dilution, cleaning of sample flow components, sample preparation, filling of the impedance counting detection unit 500, and optical detection of the sheath pushing fluid.
  • the fluid power device 900 only has two highly reusable syringes, which can reduce the cost and volume of the instrument.
  • the first syringe 910 and the second syringe 920 are driven by independent power mechanisms respectively.
  • the fluid power device 900 further includes a first motor (not shown) for driving the first syringe 910 and a second motor (not shown) for driving the second syringe 920.
  • the first motor and the third The two motors are two motors set independently of each other.
  • the step size of the second motor is preferably larger than the step size of the first motor.
  • the optical detection unit 300 includes a flow chamber 310 and an optical detection element 320.
  • the flow chamber 310 has a detection area, a diluent inlet d, a sample inlet e and a first outlet f.
  • the diluent inlet d, the sample inlet e and the first outlet f are respectively connected with The detection area is connected.
  • the detection area is used for the optical detection sample to pass through under the diluent.
  • the optical detection element 320 may include a light source, a forward scattered light signal collection device disposed on the optical axis, a side scattered light signal collection device disposed on the side of the optical axis, and a fluorescence signal collection device.
  • the optical channel mixing chamber 200 is used to provide a reaction place for the sample to be tested and the reagent to prepare the optical sample to be tested.
  • the optical channel mixing chamber 200 includes a first mixing chamber 210 and a second mixing chamber 220 .
  • the first mixing chamber 210 and the second mixing chamber 220 are two mutually independent mixing chambers, that is, the first mixing chamber 210 and the second mixing chamber 220 are two independent mixing chambers. Quantity is two.
  • the first mixing chamber 210 is used to provide a reaction place for, for example, the first sub-sample and the detection reagent (such as the first and second reagents, or the first, second and third reagents) to prepare the first test reagent. Sample.
  • the second mixing chamber 220 is used to provide a reaction place for the second subsample and detection reagents (such as the third and fourth reagents) to prepare the second sample to be tested.
  • detection reagents such as the third and fourth reagents
  • two sub-samples are prepared in two mutually independent mixing chambers as an example for description.
  • the blood cell analyzer in some embodiments of the present disclosure may also include a third mixing chamber for processing the third subsample. Analyzers with multiple mixing chambers can, on the one hand, avoid the problem of cross-contamination due to reagent residues when sharing a mixing chamber; on the other hand, they can prepare multiple subsamples at the same time, which is beneficial to improving detection efficiency.
  • the transport pipeline assembly 800 includes an optical sample preparation pipeline 810, a sampling transport pipeline 820, a first connecting pipeline 830, a second connecting pipeline 840, a third connecting pipeline 850, a fourth connecting pipeline 860, and a fifth connecting pipeline.
  • the optical sample preparation pipeline 810 is connected to the sample inlet e, the optical channel mixing chamber 200, the first syringe 910, and the second syringe 920 respectively, and the first diluent delivery pipeline 8120 is connected to the diluent inlet d and the diluent supply device respectively.
  • 1100 is connected
  • the second diluent transport pipeline 8200 is connected to the diluent supply device 1100, the hemoglobin detection unit 400, and the second syringe 920 respectively
  • the third diluent transport pipeline 8140 is connected to the diluent supply device 1100 and the impedance counting detection unit respectively.
  • the second syringe 920 is connected.
  • the diluent supply device 1100 is used to provide diluent.
  • the second syringe 920 is respectively connected to the diluent supply device 1100 and the diluent inlet d through the first diluent delivery pipeline 8120 for driving the diluent to be delivered from the diluent supply device 1100 to the flow chamber 310; the second syringe 920 is connected to the optical
  • the sample preparation pipeline 810 is connected to drive the optical detection sample to be transported from the optical channel mixing chamber 200 to the optical sample preparation pipeline 810; the second injection
  • the second syringe 920 is also connected to the diluent supply device 1100 and the hemoglobin detection unit 400 through the second diluent delivery pipeline 8200, for driving the diluent to be delivered from the diluent supply device 1100 to the hemoglobin detection unit 400; the second syringe 920 also passes through
  • the third diluent transport pipeline 8140 is respectively connected
  • the sample suction device 100 is used to collect samples to be tested.
  • the sample to be tested is a blood sample.
  • the sample to be tested can also be a body fluid sample.
  • the sampling delivery tube 820 is connected between the first syringe 910 and the sample suction device 100 .
  • the first syringe 910 is connected to the sample suction device 100 for driving the sample suction device 100 to suck the sample to be tested (ie, sampling) and driving the sample to be tested in the sample suction device 100 to be transported to the hemoglobin detection unit 400 and the optical channel mixing chamber respectively.
  • the reaction will take place within 200 (i.e. blood separation).
  • the sample suction device 100 may be a sampling needle or a sampling straw.
  • the blood cell analyzer also includes a power component (not shown) for driving the movement of the sample suction device 100.
  • the sample suction device 100 can be driven by the power component to move to a sample storage container (such as a test tube, etc.) for sampling, and then the sample suction device 100 is driven by the power component. They move to the optical channel mixing chamber 200 and the hemoglobin detection unit 400 respectively under the driving of , and perform blood separation under the driving of the first syringe 910 .
  • the first syringe 910 is connected to the sample inlet e through the optical sample preparation pipeline 810 for driving the optical detection sample to be transported from the optical sample preparation pipeline 810 to the flow chamber 310 .
  • the second syringe 920 is used to drive the first sample to be tested from the first mixing chamber 210 to the optical sample preparation pipeline 810 and drive the diluent to be transported into the flow chamber 310; the first syringe 910 is used for The first sample to be tested is driven to be transported from the optical sample preparation pipeline 810 to the flow chamber 310; the optical detection element 320 is used to perform original cell detection on the first sample to be tested entrained by the diluent through the detection area, and The original cell detection results are obtained based on the scattered light signal and antibody fluorescence signal.
  • the first syringe 910 is used to drive the second sample to be tested from the optical sample preparation pipeline 810 to the flow chamber 310 and drive the diluent to be transported into the flow chamber 310
  • the second syringe 920 is used to drive
  • the second sample to be tested is driven to be transported from the second mixing chamber 200 to the optical sample preparation pipeline 810;
  • the optical detection element 320 is used to perform leukocyte classification detection on the second sample to be tested entrained by the sheath fluid through the detection area.
  • the first reagent supply device 600 is used to provide reagents for the optical channel mixing chamber 200 .
  • the first reagent supply device 600 includes a first dosing pump 610 , a second dosing pump 620 , a third dosing pump 630 and a fourth dosing pump 640 .
  • the first quantitative pump 610 is connected to the first mixing chamber 210 to provide the first reagent to the first mixing chamber 210; the second quantitative pump 620 is connected to the first mixing chamber 210 to provide the first mixing chamber 210 with the first reagent.
  • Two reagents As a preferred embodiment of this embodiment, the first reagent and the second reagent respectively contain a hemolytic agent and a fluorescently labeled antibody.
  • the fluorescently labeled antibody reagent is used to enable the optical detection unit 300 to obtain antibody fluorescence when detecting original cells. Signal, hemolytic agent is mainly used to break red blood cells.
  • the first reagent and the second reagent can also be replaced by a reagent having both a hemolytic agent and a fluorescently labeled antibody.
  • a quantitative pump connected to 210, such as the first quantitative pump 610.
  • the third quantitative pump 630 is connected to the second mixing chamber 220 to provide the third reagent to the second mixing chamber 220; the fourth quantitative pump 640 is connected to the second mixing chamber 220 to provide the second mixing chamber 220 with the fourth reagent.
  • the third reagent and the fourth reagent respectively contain the second hemolytic agent and the first fluorescent dye.
  • the hemolyzing agent and the first fluorescent dye can also be included in a single reagent.
  • a quantitative pump connected to the second mixing chamber 220 There is only one, for example the third dosing pump 630 .
  • the hemoglobin detection unit 400 is used to provide a reaction site for the sample to be tested and, for example, the seventh reagent to prepare a hemoglobin detection sample, to provide a reaction site for the sample to be tested and the diluent to prepare an impedance counting detection sample, and to perform a test on the hemoglobin detection sample. Hemoglobin concentration detection.
  • the hemoglobin detection unit 400 in addition to being used as a preparation unit and a detection unit for hemoglobin detection samples, the hemoglobin detection unit 400 is also used as an impedance counting detection sample preparation unit. That is, the impedance counting detection sample preparation unit and the hemoglobin detection unit 400 are integrated.
  • the sample to be tested and the diluent can be added to the hemoglobin detection unit 400 for reaction to prepare the impedance counting detection sample, and then part of the impedance counting detection sample can be removed; and then the seventh reagent can be added to the hemoglobin detection unit 400 Prepare samples for hemoglobin testing.
  • the second reagent supply device 700 is used to provide the seventh reagent for the hemoglobin detection unit 400, and includes a fifth quantitative pump 710 connected to the hemoglobin detection unit 400.
  • the fifth reagent is, for example, a hemolytic agent capable of dissolving red blood cells in the blood sample, releasing hemoglobin in the red blood cells, and converting the hemoglobin into methemoglobin.
  • the reagents used in leukocyte classification detection and the reagents used in hemoglobin detection may also be the same hemolytic reagent, that is, the dosing pump used to add reagents to the second mixing chamber 220 and the reagent used to add reagents to the hemoglobin detection unit 400
  • the dosing pump for adding reagents is the same part.
  • the optical sample preparation pipeline 810 is disposed between the flow chamber 310 and the optical channel mixing chamber 200 .
  • the second syringe 920 is connected to the optical sample preparation pipeline 810 for driving the optical detection sample to be transported from the optical channel mixing chamber 200 to the optical sample preparation pipeline 810 .
  • the first syringe 910 is connected to the sample inlet through the optical sample preparation pipeline 810 e is used to drive the optical detection sample to be transported from the optical sample preparation pipeline 810 to the flow chamber 310 .
  • the optical sample preparation pipeline 810 includes a first sample preparation pipeline 811 and a second sample preparation pipeline 812. That is, the number of optical sample preparation pipelines 810 is two.
  • the first sample preparation pipeline 811 and the second sample preparation pipeline 812 are two mutually independent pipelines.
  • the first sample preparation pipeline 811 is connected to the first mixing chamber 210, the flow chamber 310, the first syringe 910 and the second syringe 920 respectively
  • the second sample preparation pipeline 812 is connected to the second mixing chamber 220, the flow chamber 310,
  • the second mixing chamber 220, the first syringe 910 and the second syringe 920 are connected.
  • the second syringe 920 is used to drive and transport the first sample to be tested from the first mixing chamber 210 to the first sample preparation pipeline 811 and drive the sheath liquid to be transported into the flow chamber 310.
  • a syringe 910 is used to drive the first sample to be tested from the first sample preparation pipeline 811 to the flow chamber 310 .
  • the second syringe 920 drives and transports the second sample to be tested from the second mixing chamber 220 to the second sample preparation pipeline 812 and drives the sheath liquid to transport into the flow chamber 310.
  • the first syringe 910 is used to drive the second sample to be tested to be transported from the second sample preparation pipeline 812 to the flow chamber 310 .
  • independent mixing chambers and independent sample preparation pipelines are used for primary cell detection and leukocyte classification detection respectively, which is beneficial to reducing cross-contamination.
  • the number of mixing chambers included in the optical channel mixing chamber 200 is not limited to two.
  • the first sample preparation pipeline 811 is connected to the flow chamber 310, and the other end is connected to the first mixing chamber 210 through the first connecting pipeline 830.
  • One end of the second sample preparation pipeline 812 is connected to the flow chamber 310, and the other end is connected to the flow chamber 310.
  • the third connecting pipe 850 is connected to the second mixing chamber 220.
  • the second syringe 920 is connected to the first sample preparation pipe 811 and the second sample preparation pipe 812 through the second connecting pipe 840.
  • the first syringe 910 is connected through the fourth connecting pipe 840.
  • the connecting pipe 860 is connected to the first sample preparation pipe 811 and is connected to the second sample preparation pipe 812 through the fifth connecting pipe 870 .
  • the first connecting pipeline 830 is provided with a first controllable valve 831
  • the second connecting pipeline 840 is provided with a second controllable valve 841
  • the third connecting pipeline 850 is provided with a third controllable valve 851 .
  • the second syringe 920 can drive and deliver diluent to the first mixing chamber 210; when the first control valve 8121 is closed, the second controllable valve
  • the second syringe 920 can drive the original cell detection sample to be transported from the first mixing chamber 210 to the first sample preparation pipeline 811;
  • the first control valve 8121 and the second controllable valve 841 When the second control valve 841 and the third controllable valve 851 are open, the second syringe 920 can drive and deliver the diluent to the second mixing chamber 220; when the first control valve 8121 is closed, and the second
  • the fourth connecting pipe 860 is connected to the first syringe 910 and the other end is connected to the first sample preparation pipe 811.
  • the fourth connecting pipe 860 is provided with a sixth control valve 861, and one end of the fifth connecting pipe 870 is connected to the first syringe 910 and the first sample preparation pipe 811.
  • the other end of the sixth control valve 861 is connected to the second sample preparation pipeline 812.
  • the sixth control valve 861 can control the first syringe 910 to switch and connect the first sample preparation pipeline 811 and the second sample preparation pipeline 812 respectively.
  • the fourth connecting pipe 860 and the fifth connecting pipe 870 can also be connected to the first syringe 910 respectively through two independent control valves.
  • a fifth controllable valve 862 is also provided on the fourth connecting pipeline 860.
  • the fifth controllable valve 862 is located between the first syringe 910 and the fourth controllable valve 861.
  • the sample suction device 100 The first syringe 910 is connected through a fifth controllable valve 862 .
  • One end of the sampling transport pipe 820 is connected to the sample suction device 100, and the other end is connected to the fifth controllable valve 862.
  • the fifth controllable valve 862 can control the first syringe 910 to switch and connect the sample suction device 100 and the sample preparation pipeline respectively.
  • the sample preparation pipeline and the sampling device 100 can also be connected to the first syringe 910 through two independent controllable valves.
  • the first connector 880 has a first interface a, a second interface b and a third interface c that communicate with each other.
  • the first interface a is connected to the sample inlet e of the flow chamber 310
  • the third interface c is connected to the sample inlet e of the flow chamber 310 through the second connecting pipe 840 .
  • Two syringes 920 are connected; one of the first sample preparation pipeline 811 and the second sample preparation pipeline 812 is connected between the second interface b and the optical channel mixing chamber 200, and the first sample preparation pipeline 811 and the second sample preparation pipeline 812 are connected between the second interface b and the optical channel mixing chamber 200.
  • the fourth connecting pipeline 860 is connected to the first sample through the third connector 8100.
  • the preparation pipeline 811 and the first connecting pipeline 830 are connected to the fifth connecting pipeline 870 through the fourth connector 8110 to connect the second sample preparation pipeline 812 and the third connecting pipeline 850 .
  • Both the third connector 8100 and the fourth connector 8110 are preferably three-way connectors, that is, the first syringe 910 is connected to the two optical sample preparation pipelines 810 through two three-way connectors.
  • the second connecting pipe 840 is also connected to the second syringe 920 and the diluent supply device 1100 through the first diluent delivery pipe 8120, that is, one end of the second connecting pipe 840 is connected to the first sample respectively.
  • the preparation pipeline 811 and the second sample preparation pipeline 812 are connected, and the other end of the second connection pipeline 840 is connected to the first diluent transport pipeline 8120.
  • the second syringe 920 is also used to drive the diluent from the diluent supply device 1100 to the optical sample preparation pipeline 810 and the optical channel mixing chamber 200. In this way, the second syringe 920 can clean the first sample preparation pipeline 811 for the diluent.
  • the second sample preparation line 812, the first mixing chamber 210 and the second mixing chamber 220 provide power.
  • first diluent transport pipeline 8120 One end of the first diluent transport pipeline 8120 is connected to the diluent inlet d of the flow chamber 310, and the other end is connected to the second syringe 920.
  • the diluent supply device 1100 and the second connecting pipeline 840 are respectively connected to the first diluent transport pipeline. 8120.
  • the first diluent delivery pipeline 8120 is provided with a first control valve 8121 and a second control valve 8122.
  • the first control valve 8121 is set close to the second syringe 920, and the second control valve 8122 is set close to the diluent inlet d.
  • the diluent is provided Device 1100 is connected to first control valve 8121.
  • the first control valve 8121 and the second control valve 8122 can control the opening and closing of the first diluent delivery pipeline 8120.
  • the second syringe 920 can push the sheath liquid into the flow chamber 310; when the first control valve 8121 and the second control valve 8122 are closed, the second syringe 920 can push the sheath liquid into the flow chamber 310. Syringe 920 cannot push sheath fluid into flow chamber 310 .
  • the second syringe 920 is respectively connected to the diluent supply device 1100 and the first syringe 910 through the fourth diluent delivery pipeline 8150 for driving the diluent to be delivered from the diluent supply device 1100 to the first syringe 910 .
  • the fourth diluent delivery pipeline 8150 is connected between the first diluent delivery pipeline 8120 and the first syringe 910 .
  • the fourth diluent delivery pipeline 8150 is provided with a fifth control valve 8151.
  • the second syringe 920 can drive the diluent to deliver the diluent to the first syringe 910 to facilitate cleaning of the second diluent.
  • the impedance counting detection unit 500 is used to perform impedance counting detection on the impedance counting detection sample.
  • the impedance counting detection may include red blood cell number detection and/or platelet count detection.
  • the detection sample for impedance counting detection can be prepared by reacting the sample to be tested and the diluent.
  • the impedance counting and detecting unit 500 includes an impedance counting and detecting device 510, a diluent storage tank 520, a positive pressure power source 530, a pressure sensor 540 and a fifth diluent transport pipeline 550.
  • the fifth diluent transport pipeline 550 is provided with a third Six control valves 551, a positive pressure power source 530, a pressure sensor 540 and a diluent reservoir 520 respectively.
  • the positive pressure power source 530 is connected to the diluent reservoir 520 through an air valve 570.
  • the diluent storage tank 520 is connected to the impedance counting detection device 510 through the fifth diluent delivery pipeline 550, and is connected to the second syringe 920 and the diluent supply device 1100 through the third dilution delivery pipeline 8140; the impedance counting sample preparation pipeline One end of the 8170 is connected to the impedance meter The detection device 510 is connected to the second syringe 920 through the first delivery pipeline 8160. The other end of the impedance counting sample preparation pipeline 8170 is connected to the hemoglobin detection unit 400 through the second delivery pipeline 8180 and connected to the third delivery pipeline 8210.
  • the second syringe 920 is also used to drive the diluent to be transported from the diluent supply device 1100 to the diluent reservoir 520 and to drive the impedance counting detection sample to be transported from the hemoglobin detection unit 400 to the impedance counting sample preparation pipeline 8170
  • the positive pressure power source 530 is used to drive the diluent from the diluent reservoir 520 to the impedance counting detection device 520, so that the impedance counting detection sample flows in the impedance counting detection device 520 under the entrapment of the diluent
  • the first syringe 910 is also used to drive the impedance counting detection sample to be transported from the impedance counting sample preparation pipeline 8170 to the impedance counting detection device 510 .
  • one end of the third diluent transport pipeline 8140 is connected to the first diluent transport pipeline 8120, and the other end is connected to the diluent reservoir 520; one end of the first transport pipeline 8160 is connected to the third diluent transport pipeline. The other end is connected to the impedance counting sample preparation pipeline 8170 and the impedance counting detection device 510 respectively.
  • the first delivery pipeline 8160 is provided with a seventh control valve 8161
  • the third diluent delivery pipeline 8140 is provided with a fourth control valve 8141
  • the fifth diluent delivery pipeline 550 is provided with a sixth control valve 551.
  • the second syringe 920 can drive the diluent to transport the diluent reservoir 520 to achieve filling; when the air valve 570 and the sixth control valve 551 are opened, The pressure power source 530 can drive the sheath liquid to be transported from the diluent reservoir 520 to the impedance counting detection device 510 .
  • the third diluent transport pipeline 8140 indirectly connects the diluent supply device 1100 and the second syringe 920 through the first diluent transport pipeline 8120; of course, in specific applications, as an alternative embodiment, It can be changed to a solution in which the third diluent delivery pipeline 8140 directly connects the diluent supply device 1100 and the second syringe 920 .
  • One end of the impedance counting sample preparation pipeline 8170 is connected to the impedance counting detection device 510 and the first transport pipeline 8160 through the seventh joint 8190, and the other end of the impedance counting sample preparation pipeline 8170 is connected to the second transport pipeline 8180 and the third transport pipeline 8160 respectively. Delivery line 8210.
  • the second transport pipeline 8180 is connected between the impedance counting sample preparation pipeline 8170 and the hemoglobin detection unit 400 .
  • An eighth control valve 8181 is provided on the second delivery pipeline 8180. When the seventh control valve 8161 and the eighth control valve 8181 are opened, the second syringe 920 can drive the impedance counting detection sample to be transported from the hemoglobin detection unit 400 into the impedance counting sample preparation pipeline 8170.
  • the impedance counting detection device 510 has a front tank (not shown in the figure), a rear tank (not shown in the figure), a gem hole (not shown in the figure), a second inlet h, a first inlet i and a second outlet g.
  • the second inlet h The first entrance i is connected to the front pool respectively, the gem hole is used to connect the front pool and the rear pool, and the second outlet g is connected to the rear pool.
  • the second inlet h is connected to the fifth diluent transport pipeline 550, the first inlet i is connected to the first transport pipeline 8160 and the impedance counting sample preparation pipeline 8170 through the seventh connector 8190, and the second outlet g is connected to the waste liquid pool 1000. connect.
  • the second diluent transport pipeline 8200 is connected between the first diluent transport pipeline 8120 and the hemoglobin detection unit 400.
  • the impedance counting sample preparation pipeline 8170 is connected to one end of the third transport pipeline 8210 and the second transport pipeline 8180 respectively.
  • One end of the third delivery pipeline 8210 is connected to the first syringe 910
  • the other end of the second delivery pipeline 8180 is connected to the second diluent delivery pipeline 8200 .
  • the second diluent delivery pipeline 8200 is provided with a third control valve 8201
  • the fifth dosing pump 710 is connected to the second diluent delivery pipeline 8200
  • the fifth dosing pump 710 and the second delivery pipeline 8180 are connected to the second dilution liquid delivery pipeline 8200.
  • connection points on the liquid delivery pipeline 8200 are all located between the third control valve 8201 and the hemoglobin detection unit 400 .
  • the second syringe 920 can deliver the diluent to the hemoglobin detection unit 400 .
  • the second diluent transport pipeline 8200 indirectly connects the diluent supply device 1100 and the second syringe 920 through the first diluent transport pipeline 8120; of course, in specific applications, as an alternative embodiment, It may be changed to a solution in which the second diluent delivery pipeline 8200 directly connects the diluent supply device 1100 and the second syringe 920 .
  • the third delivery pipeline 8210 is provided with a ninth control valve 8211, and the fifth controllable valve 862 is located between the ninth control valve 8211 and the first syringe 910.
  • the fourth connecting pipe 860 is connected to the ninth control valve 8211.
  • the fifth controllable valve 862 can control the opening and closing of the channel between the first syringe 910 and the sampling delivery pipe 820 and the opening and closing of the channel between the first syringe 910 and the ninth control valve 8211, that is, by regulating the fifth controllable valve.
  • first syringe 910 The state of communicating with the sampling delivery tube 820 and switching the state of the first syringe 910 communicating with the sample preparation pipeline (including the optical sample preparation pipeline 810 and the impedance counting sample preparation pipeline 8170).
  • the ninth control valve 8211 and the fifth controllable valve 862 Through the cooperation of the ninth control valve 8211 and the fifth controllable valve 862, the opening and closing of the channel between the first syringe 910 and the impedance counting sample preparation pipeline 8170 can be controlled; through the sixth control valve 861, the ninth control valve 8211 and The cooperation of the fifth controllable valve 862 can control the opening and closing of the channel between the first syringe 910 and the first sample preparation pipeline 811 and the second sample preparation pipeline 812 .
  • the first syringe 910 is connected to the impedance counting sample preparation pipeline 8170 and the optical sample preparation pipeline 810 through the ninth control valve 8211 respectively.
  • the optical sample preparation pipeline 810 and the impedance counting sample preparation pipeline 8170 can also be connected to the first syringe 910 through independent control valves.
  • the first drain pipe 8220 is connected between the first outlet f of the flow chamber 310 and the waste liquid pool 1000.
  • the first drain pipe 8220 is provided with a first drain control valve 8221.
  • the first drain control valve 8221 can be opened to transport the reticulocyte detection sample and diluent or the leukocyte classification detection sample and diluent flowing out from the flow chamber 310 through the detection area to the waste. Liquid pool 1000.
  • the second drain pipe 8230 is connected between the first mixing chamber 210 and the waste liquid pool 1000.
  • the second drain pipe 8230 is provided with a second drain control valve 8231. By opening the second drain control valve 8231, the first mixing chamber 210 can be emptied, so that when the first mixing chamber 210 is cleaned, the cleaning waste liquid can be discharged to the waste liquid pool 1000.
  • the third drain pipe 8240 is connected between the second mixing chamber 220 and the waste liquid pool 1000.
  • the third drain pipe 8240 is provided with a third drain control valve 8241. By opening the third drain control valve 8241, the second mixing chamber 220 can be emptied, so that when the second mixing chamber 220 is cleaned, the cleaning waste liquid can be discharged to the waste liquid pool 1000.
  • the fourth drain pipe 8250 is connected between the hemoglobin detection unit 400 and the waste liquid pool 1000.
  • the fourth drain pipe 8250 is provided with a fourth drain control valve 8251. By opening the fourth drain control valve 8251, the hemoglobin detection unit 400 can be emptied, so that when the hemoglobin detection unit 400 is cleaned, the cleaning waste liquid can be discharged to the waste liquid pool 1000.
  • the fifth drain pipe 8260 is connected between the second outlet g of the impedance counting detection device 510 and the waste liquid pool 1000 .
  • the impedance counting detection sample and the sheath fluid flowing out from the second outlet g of the impedance counting detection device 510 may be discharged to the waste liquid pool 1000 .
  • waste liquid pool 1000 there may be only one waste liquid pool 1000, namely the first liquid drain pipe 8220, the second liquid drain pipe 8230, the third liquid drain pipe 8240, the fourth liquid drain pipe 8250 and the fifth liquid drain pipe.
  • the pipeline 8260 is connected to the same waste liquid pool 1000; of course, as an alternative embodiment, the waste liquid pool 1000 can also be provided with more than two, a first liquid drain pipe 8220, a second liquid drain pipe 8230, and a second liquid drain pipe 8230. At least two of the three drain lines 8240 , the fourth drain line 8250 and the fifth drain line 8260 are respectively connected to different waste liquid pools 1000 .
  • sample analyzer 2100 according to another specific embodiment of the present disclosure with reference to FIGS. 17 and 18 .
  • the sample analyzer 2100 provided in this embodiment focuses on the implementation of an integrated machine for primary cell detection and routine blood detection.
  • the sample analyzer 2100 includes a sample distribution device 2111, an optical detection device 2112, a first reaction device 2113, a second reaction device 2116, a first sample transport component 2114 and a controller 2120; sample distribution The device 2111 is used to draw a sample from the sample container 2200, and distribute at least part of the sample to the first reaction device 2113 and/or the second reaction device 2116; the first reaction device 2113 is at least used to combine the sample with the antibody reagent, and the second reaction device 2116.
  • a hemolysis reagent provides a reaction site to prepare a first test sample, in which the antibodies in the antibody reagent can combine with the surface antigens of the original cells in the sample; the second reaction device 2116 is at least used to combine the sample with the first type of reagent A reaction site is provided to prepare and form a second sample to be tested; the first sample transport component 2114 is used to transport the first sample to be tested from the first reaction device 2113 to the optical detection component, and is used to transport the second sample to be tested The sample is transported from the second reaction device 2116 to the optical detection component; the optical detection component includes a flow chamber, a light emitting component and a light receiving component.
  • the flow chamber is used to entrap the first sample to be tested or the second sample to be tested in the sheath liquid.
  • the light-emitting component is used to emit light toward the first sample to be tested or the second sample to be tested in the flow chamber, and the light-receiving component is used to receive the light emitted by the light-emitting component and generated by the first sample to be tested.
  • the controller 2120 is configured to: analyze and obtain the first particle information in the first sample to be tested based on the first optical signal fed back by the light receiving component , the first particle information includes original cell information; according to the second optical signal fed back by the light receiving component, the second particle information in the second test sample is analyzed, and the second particle information includes neutrophil information, eosinophils Cell information, monocyte information and lymphocyte information.
  • independent places are set up for the classification of primitive cells and leukocytes, which is beneficial to allowing the reactions in the detection of leukocytes and the reactions in the detection of primitive cells to be carried out simultaneously, thereby improving the detection throughput.
  • White blood cell classification and counting and raw cell counting and detection share the same optical detection device 2112, which is low cost.
  • since manual distribution of samples is not required it has the advantages of high degree of automation, simple operation, saving time and effort, and high efficiency, and eliminates the hidden danger of human quantitative errors.
  • the sample analyzer 2100 is provided with a fixed first reaction device 2113 to carry out the reaction between the sample and the antibody reagent in the original cell detection, and there is no need to set up a blank sample container 2200 outside the sample analyzer 2100 for the reaction between the sample and the antibody reagent. , reducing unnecessary consumption of the sample container 2200.
  • the sample analyzer 2100 further includes a housing 2130, in which the optical detection device 2112, the first reaction device 2113 and the second reaction device 2116 are located.
  • the antibody reaction for primary cell detection is performed inside the sample analyzer 2100. There is no need to set up a blank sample container 2200 outside the sample analyzer 2100 for the reaction between the sample and the antibody reagent, which reduces unnecessary consumption of the sample container 2200. .
  • the sample analyzer 2100 further includes a reagent storage device 2115.
  • the reagent storage device 2115 is used to store at least a first reagent container.
  • the first reagent container is used to load antibody reagents.
  • the reagent storage device 2115 is provided in the housing 2130.
  • the antibody reagent is also stored in the sample analyzer 2100.
  • the controller 2120 is further configured to: during the sample distribution process of a single original cell detection item, control the sample volume distributed by the sample distribution device 2111 to the first reaction device 2113 to be less than or equal to 60 ⁇ L.
  • control the sample volume distributed by the sample distribution device 2111 to the first reaction device 2113 to be less than or equal to 60 ⁇ L.
  • the controller 2120 is further configured to: during the sample distribution process of a single original cell detection item, control the sample amount distributed by the sample distribution device 2111 to the first reaction device 2113 to be between 50 ⁇ L ⁇ 10 ⁇ L. It is understandable that if the sample size used for testing is too large, the amount of samples that need to be collected from the patient will be relatively large; and if the sample size used for testing is too small, it will not be conducive to ensuring the accuracy of the test results. In this embodiment, the sample volume used for a single primitive cell detection project is controlled to about 50 ⁇ L, which is beneficial to reducing the sample volume and ensuring the accuracy of the primitive cell detection results.
  • the controller 2120 is further configured to: during the sample distribution process of a single original cell detection item, control the sample volume distributed by the sample distribution device 2111 to the first reaction device 2113 to be 50uL.
  • the sample volume used for a single original cell detection project is controlled at 50 ⁇ L.
  • the controller 2120 is further configured to: during the dispensing process of the antibody reagent for a single original cell detection item, control the amount of the antibody reagent distributed to the first reaction device 2113 to be between 5 ⁇ L ⁇ 2 ⁇ L.
  • the controller 2120 is further configured to: during the dispensing process of the antibody reagent for a single original cell detection item, control the amount of the antibody reagent distributed to the first reaction device 2113 to be less than or equal to 5 ⁇ L.
  • control the amount of the antibody reagent distributed to the first reaction device 2113 to be less than or equal to 5 ⁇ L.
  • the controller 2120 is further configured to: during the dispensing process of the antibody reagent for a single original cell detection item, control the amount of the antibody reagent distributed to the first reaction device 2113 to be less than 5 ⁇ L.
  • the controller 2120 is further configured to: during the detection process of a single original cell detection item, control the reaction time between the sample and the antibody reagent in the first reaction device 2113 to be less than or equal to 90 seconds.
  • This implementation optimizes the design of the preparation method of the test sample for original cell detection and optimizes the design of the analysis method of the original cell detection results, thereby reducing the number of samples and antibody reagents while ensuring the accuracy of the test results. reaction time, thereby improving the detection efficiency of original cells.
  • the controller 2120 is further configured to: during the detection process of a single original cell detection item, control the reaction time between the sample and the antibody reagent in the first reaction device 2113 to be less than or equal to 60 seconds.
  • the antibody reagent includes a fluorescently labeled antibody that can bind to the original cell surface antigen, and the antibody is labeled with a fluorescent dye.
  • the first optical signal includes a first side scattered light signal generated by the light emitted by the light emitting component irradiating the first sample to be tested and a first side scattered light signal generated by the fluorescent labeling of the light emitted by the light emitting component irradiating the first sample to be tested.
  • First fluorescent signal The first side scattered light signal and the first fluorescence signal can distinguish primitive cells from mature leukocytes.
  • this embodiment utilizes the different binding degrees of fluorescently labeled antibodies to different cell surface antigens to obtain different fluorescent signals.
  • the side scattered light that reflects the complexity of the internal structure of the cell, it can directly distinguish the original cells from the original cells. other white blood cells, thus facilitating accurate calculation of original cells.
  • the antibody reagent is a CD45 reagent.
  • the CD45 reagent includes a fluorescently labeled antibody that can bind to the surface antigen of the original cell, and the antibody can specifically bind to the surface antigen of the original cell to facilitate the detection of the disease through fluorescence.
  • the signal differentiates blasts from white blood cells.
  • the fluorescent dye and the antibody are integrated, that is, the fluorescent reagent and the antibody are combined into one reagent, and the first reaction solution is made by reacting the sample with a single reagent.
  • the fluorescent dye and the antibody can also be separated into two different reagents, that is, the reagent used to react with the sample to form the first reaction solution can also be two reagents, one One is an antibody reagent, and the other is a fluorescent reagent.
  • the antibody reagent contains an antibody that is not fluorescently labeled but can bind to the original cell surface antigen
  • the first reaction device 2113 is used to provide a reaction site for the sample, the antibody reagent, and the first fluorescent reagent to prepare
  • the first reaction liquid is formed, and the first optical signal includes a first side scattered light signal generated by the light emitted by the light emitting component irradiating the first sample to be tested and the light emitted by the light emitting component irradiating the first sample to be tested.
  • the first fluorescent signal is generated by the first fluorescent reagent.
  • the first reaction device 2113 is used to provide a reaction site for the first reaction solution, the first hemolysis reagent, and the second fluorescent reagent to prepare and form the first sample to be tested; the first optical signal also includes light emission. The light emitted by the component irradiates the first sample to be tested and generates a second fluorescent signal via the second fluorescent reagent.
  • the controller 2120 analyzes and obtains the first particle information in the first test sample according to the first optical signal fed back by the light receiving component: analyzes and obtains the first original cell information and The first blood shadow area information is analyzed according to the second fluorescence signal to obtain the second blood shadow area information; the first original cell information and the first blood shadow area information are deducted from the second blood shadow area information to obtain the first to-be-tested Such original cell information.
  • a second fluorescent reagent in the hemolysis reaction it is beneficial to better mark the blood shadow area; by deducting the impact of the blood shadow area on the detection of original cells, more accurate original cell information can be obtained.
  • the controller 2120 can also obtain mature leukocytes that are distinguished from original cells. information.
  • the sample distribution device 2111 is also used to absorb the antibody reagent from the first reagent container, and distribute the absorbed antibody reagent to the first reaction device 2113.
  • the sample distribution device 2111 is used for time-sharing and multiplexing of sample distribution and antibody reagent distribution, which can achieve the effect of not requiring manual distribution of antibody reagents without adding an additional distribution device, which is conducive to further improving sample analysis.
  • the degree of automation of the instrument 2100 eliminates the hidden danger of errors in manual quantitative antibody reagents.
  • the antibody reagent is not limited to being distributed by the sample distribution device 2111.
  • the sample analyzer 2100 also includes a third A reagent distribution device.
  • the first reagent distribution device is used to absorb antibody reagents from the first reagent container and distribute the absorbed antibody reagents to the first reaction device 2113.
  • a third reaction device different from the sample distribution device 2111 is used.
  • a reagent dispensing device distributes antibody reagents.
  • the second reaction device 2116 includes a third reaction pool and at least one of a fourth reaction pool and a fifth reaction pool.
  • the third reaction pool is used to provide a reaction field for the sample and the first type of reagent. Therefore, the second sample to be tested is prepared, the fourth reaction pool is used to provide a reaction place for the sample and the second type of reagent to prepare the third sample to be tested, and the fifth reaction pool is used to provide a reaction place for the sample and the third type of reagent.
  • the reaction site is thus prepared to form a fourth sample to be tested.
  • the second test sample is used for four-class counting of white blood cells, that is: the third reaction pool is the DIFF reaction channel, and the second test sample is used to detect neutrophil information, eosinophil information, and monocyte information in the sample. Cell information and lymphocyte information.
  • the third sample to be tested is used for counting basophils in white blood cell classification, that is, the fourth reaction pool is the WNB reaction channel, and the third sample to be tested is used to detect basophil information in the sample. Combining the test results of the second test sample and the third test sample, the five-category counting of white blood cells is achieved.
  • the fourth sample to be tested is used for counting reticulocytes, that is, the fifth reaction pool is the RET reaction channel, and the fourth sample to be tested is used to detect reticulocyte information in the sample.
  • the second reaction device 2116 includes a third reaction pool, a fourth reaction pool and a fifth reaction pool.
  • the sample analyzer 2100 can also perform white blood cell classification counting and reticulocyte counting.
  • the arrangement of the second reaction device 2116 is not limited to this.
  • the second reaction device 2116 can also be used only for white blood cell classification and counting, and the white blood cell classification and counting can either be Five-category counting can also be four-category counting, or three-category counting; or, as another alternative embodiment, the second reaction device 116 can also include only one reaction pool, which can be used for leukocyte differential counting. It can also be used for the preparation of test samples for reticulocyte counting.
  • the first sample transport assembly 2114 is also used to transport the second sample to be tested, the third sample to be tested, or the fourth sample to be tested from the second reaction device 2116 to the flow chamber.
  • the second reaction device 2116 includes a third reaction pool, a fourth reaction pool and a fifth reaction pool
  • the first sample transport component 2114 is time-shared and multiplexed to: transport the first sample to be tested from the first reaction pool 21131 to the flow chamber, transport the second sample to be tested from the third reaction tank to the flow chamber, transport the third sample to be tested from the fourth reaction tank to the flow chamber, transport the fourth sample to be tested from the fifth reaction tank Transported to flow chamber.
  • the original cell channel and the blood routine optical detection channel share a sample transport component to transport the test sample to the flow chamber, which has the advantages of simple structure and low cost; of course, in specific applications, as an alternative embodiment , the original cell channel and the blood routine optical detection channel can also use different sample transport components to transport the test sample to the flow chamber.
  • the flow chamber is also used to allow the second sample to be tested, the third sample to be tested, or the fourth sample to be tested to pass under the entrapment of the sheath liquid.
  • the second reaction device 2116 includes a third reaction pool, a fourth reaction pool, and a fifth reaction pool
  • the flow chamber is multiplexed in a time-sharing manner: for the first sample to be tested to pass through under the entrapment of the sheath liquid, and for the second sample to be tested to pass through.
  • the test sample passes through the sheath liquid
  • the third sample to be tested passes through the sheath liquid
  • the fourth sample to be tested passes through the sheath liquid.
  • the light emitting component is also used to emit light toward the second sample to be tested, the third sample to be tested, or the fourth sample to be tested in the flow chamber.
  • the second reaction device 2116 includes a third reaction cell, a fourth reaction cell and a fifth reaction cell
  • the light emitting component is time-shared and multiplexed: emitting light toward the first sample to be tested in the flow chamber, toward the flow chamber.
  • the second sample to be tested emits light toward the third sample to be tested in the flow chamber and the light toward the fourth sample to be tested in the flow chamber.
  • the light receiving component is also used to receive the second optical signal generated by the light emitted by the light emitting component via the second test sample, or the third optical signal generated by the third test sample, or the second optical signal generated by the third test sample.
  • the second reaction device 2116 includes a third reaction cell, a fourth reaction cell and a fifth reaction cell
  • the light receiving component is time-shared and multiplexed to: receive the light emitted by the light emitting component and generate the first test sample through the first test sample.
  • the optical signal is a second optical signal generated by receiving the light emitted by the light-emitting component and passing through the second sample to be tested, or the third optical signal generated by the third sample to be tested, or the third optical signal generated by the fourth sample to be tested.
  • the first optical signal includes a first side scattered light signal generated by irradiating light emitted by the light emitting component to the first sample to be tested, and irradiating the light emitted by the light emitting component to the first sample to be tested.
  • the first fluorescent signal generated by the fluorescent label or the first fluorescent reagent on the sample and the light emitted by the light emitting component are irradiated to the second fluorescent signal generated by the second fluorescent reagent on the first sample to be tested.
  • the first type of reagent includes a third fluorescent reagent and a second hemolytic reagent
  • the second optical signal includes a second side scattered light signal generated by irradiating light emitted by the light emitting component onto the second sample to be tested. and a third fluorescent signal.
  • the second type of reagent includes a fourth fluorescent reagent and a third hemolytic reagent
  • the third optical signal includes a third side scattered light signal generated by irradiating light emitted by the light emitting component onto the third sample to be tested. and a fourth fluorescent signal.
  • the third type of reagent includes a fifth fluorescent reagent
  • the fourth optical signal includes a fourth side scattered light signal and a fifth fluorescent signal generated by irradiating light emitted by the light emitting component onto the fourth sample to be tested.
  • the light receiving component includes a side scattered light detector, a forward scattered light detector, a first fluorescence detector and a second fluorescence detector
  • the side scattered light detector is used to receive the first side scattered light signal, the second side scattered light signal, the third side scattered light signal and the fourth side scattered light signal.
  • the forward scattered light detector is used to receive the light emitted by the light emitting component and illuminate the front light generated by the sample to be tested.
  • the first fluorescence detector is used to receive the first fluorescence signal
  • the second fluorescence detector is used to receive the second fluorescence signal, the third fluorescence signal, the fourth fluorescence signal and the fifth fluorescence signal.
  • the controller 2120 is further configured to perform at least one analysis action as follows based on the feedback information of the light-receiving component: based on the second optical signal fed back by the light-receiving component, analyze and obtain the third component in the second sample to be tested.
  • Two particle information, the second particle information includes neutrophil information, eosinophil information, monocyte information and lymphocyte information; according to the third optical signal fed back by the light receiving component, the third sample to be tested is analyzed and The third particle information, the third particle information at least includes basophil information; according to the fourth optical signal fed back by the light receiving component, the fourth particle information in the fourth test sample is analyzed, and the fourth particle information includes Reticulocyte information.
  • the controller 2120 is time-shared and multiplexed to: analyze and obtain the original cell count information and leukocyte count information of the sample based on the information fed back by the light receiving component. Classification count information and reticulocyte count information.
  • the second reaction device 2116 at least includes a third reaction pool
  • the controller 2120 is further configured to: deduct the abnormal lymphocyte data in the second particle information from the original cell information of the first particle information, Obtain measurement data of original cells. Abnormal lymphocytes will affect the identification of particles in the original sensitive area. The data of abnormal lymphocytes are measured through the DIFF detection channel, and the data measured on the original cell channel are deducted from the abnormal lymphocyte data measured on the DIFF detection channel, thereby obtaining a more accurate raw cell data.
  • the second reaction device 2116 at least includes a fourth reaction pool
  • the controller 2120 is further configured to: deduct basophils in the third particle information from the original cell information of the first particle information. data to obtain the measurement data of the original cells. Basophils will affect the identification of particles in the original sensitive area. The data of basophils are measured through the WNB detection channel, and the data measured in the original cell channel are deducted from the basophils measured by the WNB detection channel. data to obtain more accurate original cell data.
  • the controller 2120 is configured to control the sample distribution device 2111 to perform the following actions in sequence: draw the sample from the sample container 2200, and draw the sample from the sample container 2200. Absorb the antibody reagent from a reagent container, distribute the absorbed antibody reagent and part of the sample to the first reaction device 2113, and distribute the absorbed part of the sample to the second reaction device 2116, where the sample distribution device 2111 draws from the sample container 2200 at one time
  • the sample volume is greater than or equal to the sum of the sample volume distributed to the first reaction device 2113 and the sample volume distributed to the second reaction device 2116 .
  • the sample distribution device 2111 draws the original cell testing items and routine blood testing items from the sample container 2200 at one time. sample size, and then draw the The fluorescently labeled antibody reagent distributes the antibody reagent and the sample of the original cell detection item to the original cell reaction channel, and then allocates the sample of the routine blood test item to the routine blood reaction channel. Since the reaction time of the original cell test is longer than that of the routine blood test, the samples of the original cell reaction channel are allocated first, and then the samples of the blood routine test channel are allocated, which will help shorten the test result output time of the same sample.
  • the sample dividing sequence of the sample analyzer 2100 is not limited to the above method.
  • the controller 2120 is configured to control the sample distribution device 2111 to perform the following actions in sequence: from The sample is sucked into the sample container 2200, and part of the sample is distributed to the second reaction device 2116.
  • the antibody reagent is sucked from the first reagent container, and the absorbed antibody reagent and part of the sample are distributed to the first reaction device 2113, where the sample is distributed
  • the amount of sample that the device 2111 draws from the sample container 2200 at one time is greater than or equal to the sum of the amount of the sample assigned to the first reaction device 2113 and the amount of the sample assigned to the second reaction device 2116 .
  • the sample is aspirated once, and the routine blood test items are first divided into samples, and then the antibody reagent is drawn, and then the original cell test items are divided into antibody reagents and samples.
  • the controller 2120 is configured to control the sample distribution device 2111 to perform the following actions in sequence: from Draw the first sample from the sample container 2200, draw the antibody reagent from the first reagent container, distribute the drawn antibody reagent and at least part of the first sample to the first reaction device 2113, and draw the second sample from the sample container 2200 , distribute at least part of the aspirated second sample to the second reaction device 2116.
  • the original cell test and the routine blood test are aspirated separately. First, the original cell test item is aspirated, the antibody reagent is aspirated, the antibody reagent is separated, and the sample is divided, and then the blood routine test item is aspirated and sampled. Sample.
  • the controller 2120 is configured to control the sample distribution device 2111 to perform the following actions in sequence: from The first sample is drawn from the sample container 2200, and at least part of the drawn first sample is distributed to the second reaction device 2116.
  • the second sample is drawn from the sample container 2200, and the antibody reagent is drawn from the first reagent container.
  • the antibody reagent and at least part of the second sample are distributed to the first reaction device 2113.
  • the original cell test and the routine blood test are aspirated separately.
  • the routine blood test items are first aspirated and sampled, and then the original cell test items are aspirated, antibody reagents, and antibody reagents are aspirated. Sample.
  • the controller 2120 is configured to control the sample distribution device 2111 to perform the following actions in sequence: from The sample is sucked into the sample container 2200, and part of the sample is distributed to the second reaction device 2116, and part of the sample is distributed to the first reaction device 2113, and the antibody reagent is sucked from the first reagent container, and the absorbed antibody reagent is distributed to The first reaction device 2113 , wherein the sample amount drawn by the sample distribution device 2111 from the sample container 2200 at one time is greater than or equal to the sum of the sample amount distributed to the first reaction device 2113 and the sample amount distributed to the second reaction device 2116 .
  • the sample and the antibody reagent are distributed separately, and the sample is aspirated at one time, and the routine blood test items are sampled first, and then the original cell test item is sampled, and then the antibody reagent is aspirated and distributed.
  • the original cell testing items can also be sampled first, and then the routine blood testing items can be sampled, and then the antibody reagents can be aspirated and distributed.
  • the sample dispensing device 2111 draws a sample from the sample container 2200 in one go at least for distribution to the first reaction device 2113 and the second reaction device 2116, the sample dispensing device 2111 draws a sample from the sample container 2200 in one go.
  • the sample volume is less than or equal to 200 ⁇ L.
  • the sample volume for primary cell and blood routine testing is less than or equal to 200 ⁇ L, which greatly reduces the sample volume.
  • the sample analyzer 2100 further includes a hemoglobin detection device 2117.
  • the sample distribution device 2111 is also used to distribute at least part of the sample drawn to the hemoglobin detection device 2117.
  • the hemoglobin detection device 2117 is used to analyze at least the sample and the fourth The fifth test sample made of similar reagents is used to measure hemoglobin parameters.
  • the controller 2120 is also configured to: based on the information fed back by the hemoglobin detection device 2117, analyze and obtain the fifth test sample. Hemoglobin information in the sample. Hemoglobin testing is part of the routine blood testing project. Of course, hemoglobin does not necessarily need to be tested in routine blood testing.
  • the sample analyzer 2100 provided in this embodiment can also measure hemoglobin detection items, which is helpful to expand the applicable scope of the sample analyzer 2100.
  • the fourth type of reagent includes a diluent and a fourth hemolytic reagent, that is, the fifth test sample is made of a diluent and a fourth hemolytic reagent.
  • Diluent is used to dilute the sample.
  • the fourth hemolytic reagent is, for example, a hemolytic reagent capable of dissolving red blood cells in the blood sample, releasing hemoglobin in the red blood cells, and converting the hemoglobin into methemoglobin.
  • the diluent and the fourth hemolysis reagent can also be combined into one reagent.
  • the sample analyzer 2100 further includes an impedance detection device 2118.
  • the impedance detection device 2118 is used to measure a sixth test sample made of at least the sample and the fifth type of reagent.
  • the controller 2120 is also configured This is: analyzing and obtaining the platelet information and/or red blood cell information in the sixth sample to be tested based on the information fed back by the impedance detection device 2118. Platelets and red blood cells are part of routine blood tests. Of course, platelets and/or red blood cells are not necessarily detected in routine blood tests.
  • the sample analyzer 2100 provided in this embodiment can also measure platelet and/or red blood cell detection items, which is helpful to expand the applicable scope of the sample analyzer 2100.
  • the fifth type of reagent is a diluent.
  • the hemoglobin detection device 2117 includes a sixth reaction tank and a hemoglobin detection component.
  • the sixth reaction tank is at least used to provide a reaction place for the sample and the fourth type of reagent to prepare and form the fifth test sample.
  • the sixth reaction pool is the reaction channel of hemoglobin, that is, the HGB reaction channel.
  • the hemoglobin detection component is used to detect hemoglobin on the fifth test sample in the sixth reaction pool.
  • the impedance detection device 2118 includes a detection chamber and an impedance detection component.
  • the sample analyzer 2100 also includes a second sample transport component.
  • the sixth reaction cell is also used to provide a reaction place for the sample and the diluent to prepare and form the second sample.
  • the second sample transport component is connected between the sixth reaction cell and the detection chamber for transporting the sixth sample to be tested from the sixth reaction cell to the detection chamber under the control of the controller 2120 .
  • the sixth reaction cell is first used to prepare the sixth sample to be tested for impedance detection, and a part of the sixth sample to be tested is transported to the detection chamber through the second sample transport assembly for impedance detection, and then the sixth reaction cell is used for impedance detection.
  • Using at least part of the sixth test sample remaining in the sixth reaction pool to prepare the fifth test sample is equivalent to the sixth reaction pool being reused to prepare the test sample for impedance detection and the test sample for hemoglobin detection. Sample.
  • a separate reaction cell can also be set up for impedance detection to prepare test samples for impedance detection.
  • the sample dispensing device 2111 includes a sample needle 21111, a sample suction and discharge driving component, and a needle movement driving component; the sample needle 21111 is used to suck and discharge samples.
  • the sample suction and discharge driving component is used to provide driving force for sample suction and discharge.
  • the needle movement driving component is used to drive the sample needle 21111 to move in two-dimensional or three-dimensional space, so that the sample needle 21111 moves to different work stations, such as the standby position, the sample suction position, the sample addition position, and the antibody reagent suction position. , cleaning position, etc.
  • the sample suction and discharge driving component is a syringe.
  • the sample analyzer 2100 measures the sample in a sample container 2200 and outputs at least the following two test results: original cell test results and routine blood test results. If a patient needs both raw cell testing and routine blood testing, the blood samples collected from the patient can share a sample container 2200, that is, the two types of testing items can share a sample container 2200 to load samples. In this way, on the one hand, it is beneficial to reduce the consumption of the sample container 2200, and on the other hand, it is beneficial to reduce the amount of samples to be collected.
  • the samples for testing the original cell testing items and the samples for testing the routine blood testing items can also be separately packed in two different sample containers 2200.
  • the sample analyzer 2100 measures the sample drawn by the sample distribution device 2111 from the sample container 2200 at least once and outputs the following two test results: original cell test results and routine blood test results.
  • at least two detection results are outputted from one sample aspiration, which can help reduce the number of sample aspiration actions of the sample distribution device 2111, thereby improving the detection efficiency of the sample analyzer 2100.
  • samples can also be aspirated independently, that is, one sample can be The number of sampling times of the sample in the container 2200 can also be more than two times.
  • the sample analyzer 2100 further includes an identification device 2119, which is used to identify the sample container 2200.
  • the controller 2120 is also configured to obtain at least the test item information to be tested for the sample in the sample container 2200 based on the feedback information from the identification device 2119 .
  • the controller 2120 is further configured to: control the sample analyzer 2100 to perform original cell detection and/or routine blood testing on the samples in the sample container 2200 based on the test item information to be tested.
  • the sample analyzer 2100 automatically obtains the detection item information of the sample based on the feedback information from the identification device 2119, and automatically controls the analyzer execution body 2110 to execute the corresponding detection items, with a high degree of automation.
  • the sample analyzer 2100 also includes a human-computer interaction device 2140.
  • the human-computer interaction device 2140 is configured with a primitive cell detection mode and a blood routine detection mode.
  • the controller 2120 is also configured to: according to the operator's human-computer interaction
  • the detection mode selected by the device 2140 controls the sample analyzer 2100 to perform original cell detection and/or routine blood detection on the sample in the sample container 2200 .
  • the operator can manually start the sample detection mode through the human-computer interaction device 2140.
  • the human-computer interaction device 2140 includes at least one of a display screen, a keyboard, and a mouse.
  • the identification device 2119 is used to identify the identification code of the sample container 2200.
  • the identification code includes at least one of a two-dimensional code, a barcode, and a radio frequency (RFID).
  • the controller 2120 obtains the detection item information of the sample in the sample container 2200 based on the identification code recognized by the identification device 2119 .
  • the identification device 2119 can also be used to identify the appearance characteristics of the sample container 2200.
  • the appearance characteristics of the sample container 2200 include at least one of the shape of the container body, the size of the container body, the sample volume, whether there is a cap body, the color of the cap body, the shape of the cap body, and the size of the cap body.
  • the controller 2120 Based on the feedback information from the identification device 2119, the controller 2120 not only obtains the test item information of the sample, but also obtains at least one of the following information: the sample volume in the sample container 200, the type of the sample container 200, and the sample number information.
  • the identification device 2119 includes at least one of a vision camera, a barcode scanner, a QR code scanner, and a wireless radio frequency code reader.
  • Reagent A DIFF fluorescent dye (M-68P FD staining solution) 20 ⁇ L
  • Reagent B DIFF channel hemolytic reagent (M-68P LD hemolytic reagent) 1mL
  • the blood cell analyzer (Mindray BC-6800Plus, excitation wavelength 450nm) draws 20 microliters of the blood sample to be tested (taken from the subject's peripheral blood, containing immature granulocytes (IG)) through the sampling needle and sends it into the mixing chamber. Reagents A to C are added to the mixing chamber in the above amounts, mixed with the blood sample, and incubated at 42°C for 60 seconds. The test sample is prepared and detected. The particles in the test sample are collected to generate forward scattered light. Side scattered light and fluorescence signals.
  • IG immature granulocytes
  • the DIFF channel of the equipment measured the white blood cell content of the sample to be 3.4 ⁇ 10 9 /L.
  • the BLAST% was 0%, so there is no need for FL/SSC. Correction.
  • the FL/SSC scatter plot Figure 13
  • the four classification results of leukocytes were obtained as Neu%, Lym%, Mon%, Eos% and IG% were 64.2%, 7.1%, 9.2%, 4.4% and 21.1% respectively.
  • the above ratios can be further converted into the content of each particle in the sample through calculation.
  • the Neu%, Lym%, Mon%, Eos% and IG% obtained by taking the same blood sample through the classic manual microscopic counting method were 62.3%, 6.61%, 7.4%, 4%% and 18.1% respectively.
  • Reagent A DIFF fluorescent dye (M-68P FD staining solution) 20 ⁇ L
  • Reagent B DIFF channel hemolytic reagent (M-68P LD hemolytic reagent) 1mL
  • the blood sample to be tested (peripheral blood taken from the subject, containing primary cells) was processed and detected according to the same method as in Example 1 and using the same equipment.
  • the DIFF channel of the equipment measured the white blood cell (including blast cells) content of the sample to be 12.6 ⁇ 10 9 /L.
  • the BLAST% was 12%, and then the BLAST content in the sample was calculated to be 1.5 ⁇ 10 9 /L.
  • the particles are divided into BLAST, and their distribution in the FL/SSC dimension is determined (dotted line box in Figure 14). After deducting the BLAST particles in the above areas in the FL/SSC dimension, the corrected Lym and Mon areas are obtained. Particle number, the corrected classification results can be obtained: Neu%, Lym%, Mon% and Eos% are 52.9%, 33.7%, 1.1% and 0.2% respectively.
  • the BLAST%, Neu%, Lym%, Mon%, Eos% and IG% obtained by taking the same blood sample through the classic manual microscopy counting method were 10.4%, 53.2%, 31.4%, 1.2% and 0.2% respectively. It can be seen from the results that the above method has a good correlation with the proportion of blast cells and leukocytes in manual microscopy.
  • Reagent A DIFF fluorescent dye (Mindray, M-68P FD staining solution) 20 ⁇ L
  • Reagent B DIFF channel hemolytic reagent (Mindray, M-68P LD hemolytic reagent) 1mL
  • Reagent D CD45 channel hemolytic reagent (Mindray, diethylene glycol, formaldehyde, buffer) 1mL
  • Reagent E WNB fluorescent dye (M-68P FN staining solution) 20 ⁇ L
  • Reagent F WNB channel hemolytic reagent (M-68P LN hemolytic reagent) 1mL
  • the blood cell analyzer (Mindray BC-6800Plus, excitation wavelength 450nm) draws 60 microliters of the blood sample to be tested (the same blood sample as in Example 2) through the sampling needle, and divides the blood samples to be tested into three sub-blood samples into different mixing chambers. Add reagents A and B to the DIFF channel mixing chamber according to the above dosage, mix with the blood sample and incubate at 42°C for 30 seconds to prepare the second test sample; add reagents E and F to the WNB channel mixing chamber according to the above dosage.
  • the third sample to be tested is prepared; reagents C and D are added to the CD45 channel mixing chamber according to the above dosage, mixed with the blood sample and incubated at 42°C for 45 seconds and then prepared. Become the first sample to be tested.
  • the three samples to be tested are detected in sequence and the particles in the above-mentioned channel samples are collected to generate forward scattered light, side scattered light and fluorescence signals.
  • the white blood cell content of the sample measured by the DIFF channel of the equipment was 12.6 ⁇ 10 9 /L.
  • the obtained Lym% in the FL/SSC dimension of the WNB channel is 33.7% (see panel A in Figure 15).
  • the dimension of CD45FL/SSC by counting the number of scattered points in the divided BLAST region, it was found that the BLAST% was 12%, and then the original cell content in the sample was calculated to be 1.5 ⁇ 10 9 /L.
  • BLAST% (12%) is obtained according to the CD45FL/SSC dimension
  • Lym% (33.7%) is obtained from the FL/SSC dimension of the WNB channel. Combining the above two results corresponds to proportional deduction in the DIFF channel to obtain the corrected Neu%, Mon% and Eos % are 52.9%, 1.1% and 0.2% respectively.
  • Reagent D CD45 channel hemolytic reagent (Mindray, diethylene glycol, formaldehyde, buffer) 1mL
  • the blood cell analyzer (Mindray BC-6800Plus, excitation wavelength 450nm) draws 20 microliters of the blood sample to be tested (taken from the patient's peripheral blood) through the sampling needle and sends it into the mixing chamber. Reagents C and D are added to the mixing chamber according to the above dosage. After mixing with the blood sample and incubating it at 42°C for 60 seconds, a test sample is prepared and detected. The particles in the sample are collected to generate forward scattered light, side scattered light and fluorescence signals.
  • the equipment measured the white blood cell content in the sample to be 11.9 ⁇ 10 9 /L.
  • the number of scattered points in the divided BLAST, Neu, Lym, Mon and Eos regions is counted, and the results of BLAST%, Neu%, Lym%, Mon% and Eos% are respectively 11.9%, 52.9%, 33.7%, 1.1% and 0.2%.
  • the BLAST%, Neu%, Lym%, Mon%, Eos% and IG% obtained by taking the same blood sample through the classic manual microscopy counting method were 10.4%, 53.2%, 31.4%, 1.2% and 0.2% respectively. From It can be seen from the above results that the above method has a good correlation with the proportion of blast cells and leukocytes in manual microscopy.

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Abstract

检测原始细胞的血细胞分析仪及检测方法。血细胞分析仪包括：吸样装置（100）用于吸取至少部分样本作为第一子样本输送到混合室（200,210）；试剂供应装置（600,700）用于将试剂提供到混合室（200,210）；混合室（200,210）用于将第一子样本与包括第一溶血剂的第一试剂和包括经荧光标记的抗体的第二试剂混合；光源用于将光束对准与混合室（200,210）相连通的光学流动室（310）的检测孔；光学检测器用于检测通过检测孔的粒子的包括第一侧向光散射强度信号的光散射信号和包括抗体荧光强度信号的荧光信号；以及处理器与光学检测器可操作地连接，并根据第一侧向光散射强度信号和抗体荧光强度信号获得第一子样本中的粒子信息，第一子样本中的粒子信息包括原始细胞信息。

Description

检测原始细胞的血细胞分析仪及检测方法 技术领域
本公开涉及体外诊断领域，特别涉及原始细胞检测的血细胞分析仪和方法。
背景技术
健康人外周血中通常不会出现原始细胞。外周血出现原始细胞通常提示血液系统疾病，如急慢性白血病、骨髓增生异常综合征等。根据原始细胞的形态学特征，将急性白血病分为急性淋巴细胞白血病和急性髓细胞白血病两大类及其不同亚型。虽然当前临床诊断和疗效评估的金标准仍是基于骨髓检查使用MICM(morphology,immunology,cytogenetics,molecular biology)诊疗模式，但是外周血取样便捷，患者接受度高，因此检测外周血中的原始细胞仍有重要的意义。
例如，有研究表明，诊断急性髓细胞白血病(AML)时如外周血原始细胞高于2000/μL，可将外周血检查作为AML临床诊疗评估的手段。再如，诱导化疗后患者外周血原始细胞清除速率及化疗后外周血残余白血病细胞水平与患者对药物的敏感性及疗效密切相关。在AML患者首次诱导化疗过程中，可持续监测1-7天的外周血中原始细胞与初发时原始细胞的比值，当比值达到0.1时所需的天数是AML的独立预后因素。此外，另一研究报道外周血原始细胞快速清除(≤6天)能准确预测诱导化疗第14天骨髓白血病细胞清除率和诱导治疗后的完全缓解情况，并且能作为AML疗效和生存评估的良好预后因素。因此，外周血中原始细胞的检测也可以为白血病的诊疗带来较显著的价值。根据2018年ELN发布的MRD检测指南，外周血中原始细胞的检测可以实现治疗过程连续监测病情及治疗完成密切监测复发。
目前外周血原始细胞检测方法有血细胞分析仪、显微镜检法和流式细胞仪三种方法。
血细胞分析仪法目前用于临床实验室的筛查，基于国际血液学复检专家组的原始细胞报警规则，触发人工显微镜检查筛查和确认原始细胞的存在。因此，虽然血细胞分析仪具有快速、高效、操作简便等特点，但还无法精准识别和定量原始细胞，仅能给出可疑性报警，而这种报警存在较高的假阴性率以及较高的假阳性率。
显微镜检法是依靠血涂片进行显微镜人工或软件观察，是业界所认可的原始细胞分类计数的参考方法。但该方法所检测的细胞数量少，仅100-200个/人，因而检测结果的质量与水平受到标本采集、涂片、染色等的影响，在检测的精密度和原始细胞的检测灵敏度上难以显著改善。
流式细胞分析仪法是借助流式细胞技术，使用多种单克隆抗体通过白细胞等血细胞表面抗原表达的差异，对白细胞等进行分类计数。因此该方法排除了人工显微镜检查所引入的主观误差，显著增大了统计量，是一种有望成为更准确、高效的方法。但是该方法费用昂贵，操作繁琐复杂。国际血液学标准化委员会(ICSH)在2016年左右组织5家参考实验室进行流式细胞仪方法的探索，对外周血中白细胞各亚型、未成熟粒细胞、原始细胞和有核红细胞进行分类计数，需要同时用到8种单克隆抗体。而原始细胞由于细胞表面抗原无特异性，其检出依赖于排除法。
因此，需要一种操作方便，成本较低的检测外周血中原始细胞的方法。
发明内容
本公开的第一个目的在于提供一种原始细胞检测的血细胞分析仪以及原始细胞的 检测方法，该血细胞分析仪及方法可对外周血中的原始细胞进行快速且准确的分析，并能够实现计数，并且不影响对外周血样本中其他各类细胞分类及计数。
为此，本公开第一方面提供一种血液分析仪，包括吸样装置、试剂供应装置、混合室、光源、光学流动室、光学检测器、处理器和编程有计算机应用程序的非暂时性计算机可读存储介质，其中，
所述吸样装置用于吸取样本，并将所述吸取样本的至少部分作为第一子样本输送到所述混合室；
所述试剂供应装置用于提供试剂并输送到所述混合室，所述试剂包括第一类试剂；
所述混合室用于将所述第一子样本与所述第一类试剂混合以形成第一待测试样；其中所述第一类试剂包括第一试剂和第二试剂，所述第一试剂包括第一溶血剂，所述第二试剂包括经荧光标记的抗体，所述抗体能够与原始细胞的表面抗原相结合；
所述光源用于将光束对准与所述混合室相连通的所述光学流动室的检测孔；
所述光学检测器用于检测通过所述检测孔的所述第一待测试样的光散射信号和荧光信号，所述光散射信号包括来自第一待测试样的第一侧向光散射强度信号，所述荧光信号包括来自所述经荧光标记的抗体的抗体荧光强度信号；
所述处理器与所述光学检测器可操作地连接，当所述非暂时性计算机可读存储介质中的计算机应用程序被所述处理器执行时，控制所述第一溶血剂与所述第一子样本在所述混合室中的混合时间不超过2分钟，且所述处理器根据所述第一侧向光散射强度信号和所述抗体荧光强度信号获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括原始细胞信息。
根据一些实施方式，所述第一类试剂还包括第三试剂，所述第三试剂包括第一荧光染料；所述光散射信号还包括来自所述第一荧光染料的第一荧光信号；当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一侧向光散射强度信号和所述第一荧光强度信号，获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息。
根据一些实施方式，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本中不含原始细胞时，根据所述第一子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息；或者，所述处理器根据所获得的原始细胞信息，当所述原始细胞信息显示所述样本中含有原始细胞时，从所述第一子样本的粒子信息中扣除所述原始细胞的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
根据另一些实施方式，所述吸样装置还将所述吸取样本的至少部分作为第二子样本和第三子样本依次输送到所述混合室；所述试剂还包括第二类试剂，所述第二类试剂包括第三试剂、第四试剂、第五试剂和第六试剂；
所述混合室进一步用于将所述第二和第三子样本分别进行混合，其中第二子样本与第三试剂和第四试剂混合以获得第二待测试样，第三子样本与第五试剂和第六试剂混合以获得第三待测试样；其中，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同，所述第二荧光染料不同于所述第一荧光染料；
所述光学检测器进一步用于检测分别通过所述检测孔的所述第二和第三待测试样的光散射信号和荧光信号；所述光散射信号包括来自第二待测试样的第二侧向光散射强度信号、以及来自第三待测试样的第三侧向光散射强度信号；所述荧光信号包括来自第二待测试样的第一荧光染料的第二荧光强度信号和来自第三待测试样的第二荧光染料 的第三荧光强度信号；以及
当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第二待测试样的所述第二侧向光散射强度信号和所述第二荧光强度信号获得所述第二子样本中的粒子信息，和根据所述第三待测试样的所述第三侧向光散射强度信号和所述第三荧光强度信号获得所述第三子样本中的粒子信息；其中所述第二子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息，所述第三子样本中的粒子信息包括淋巴细胞信息、嗜碱性粒细胞信息和有核红细胞信息；
其中，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本不含原始细胞时，根据所述第二子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息；或者
当所述原始细胞信息显示所述样本含有原始细胞时，根据所述第二子样本中的粒子信息，结合所述原始细胞信息和所述第三子样本中的粒子信息中的淋巴细胞信息，消除所述原始细胞信息对所述第二子样本中的粒子信息的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息。
在本公开的不同实施方式中，第三试剂可包括在第一类试剂中，也可包括在第二类试剂中。
根据本公开的实施方式，当所述计算机应用程序被所述处理器执行时，使所述处理器对原始细胞进行计数。
根据本公开的实施方式，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本含有原始细胞时，报警所述样本中存在原始细胞，或报警所述样本中存在原始细胞并输出原始细胞计数值。
根据本公开的一种实施方式，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一前向光散射强度信号和所述抗体荧光强度信号确定所述原始细胞为淋系原始细胞和/或髓系原始细胞。
根据本公开的另一种实施方式，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一前向光散射强度信号和所述抗体荧光强度信号报警所述样本中存在淋系原始细胞和/或髓系原始细胞。
根据本公开的又一种实施方式，当所述计算机应用程序被所述处理器执行时，所述处理器根据所述第一侧向光散射强度信号和抗体荧光强度信号进一步获得所述样本中的白细胞信息，所述白细胞信息包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的信息。
根据本公开血液分析仪还包括血红蛋白检测单元和/或阻抗计数检测单元。
本公开第二方面提供一种样本分析方法。所述方法包括接收操作指令，对样本进行原始细胞检测。
其中，所述原始细胞检测包括以下步骤：
吸取样本，包括吸取所述样本的至少部分作为第一子样本；
处理样本，包括将所述第一子样本与第一类试剂混合以获得第一待测试样；其中所述第一类试剂包括第一试剂和第二试剂，所述第一试剂包括第一溶血剂，所述第一溶血剂与所述第一子样本的混合时间不超过2分钟，所述第二试剂包括荧光标记的抗体，所述抗体能够与原始细胞的表面抗原相结合；
检测待测试样，包括使所述第一待测试样中的粒子逐个通过光学检测装置的检测区， 并且利用所述光学检测装置的光源对所述第一待测试样中的粒子进行照射，以测得第一待测试样的光散射信号和荧光信号，所述光散射信号包括来自第一待测试样的第一侧向光散射强度信号，所述荧光信号包括来自所述荧光标记的抗体的抗体荧光强度信号；和
获取粒子信息，包括根据所述第一侧向光散射强度信号和所述抗体荧光强度信号获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括原始细胞信息。
根据一些实施方式，所述第一类试剂还包括第三试剂，所述第三试剂包括第一荧光染料，且所述光散射信号还包括来自第一待测试样中第一荧光染料的第一荧光信号；所述原始细胞检测进一步包括：根据所述第一侧向光散射强度信号和所述第一荧光强度信号，获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息。
根据一些实施方式，所述原始细胞检测进一步包括：
当所述原始细胞信息显示所述样本中不含原始细胞时，根据所述第一子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息；或者，当所述原始细胞信息显示所述样本中含有原始细胞时，从所述第一子样本中的粒子信息中扣除所述原始细胞的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
根据另一些实施方式，在所述原始细胞检测中，所述吸取样本进一步包括吸取所述样本的至少部分作为第二子样本和第三子样本；
所述处理样本进一步包括用第二类试剂处理所述第二和第三子样本，其中所述第二类试剂包括第三试剂、第四试剂、第五试剂和第六试剂，将所述第二子样本与第三试剂和第四试剂混合以获得第二待测试样，和将所述第三子样本与第五试剂和第六试剂混合以获得第三待测试样；其中，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同，以及所述第二荧光染料不同于所述第一荧光染料；
所述检测待测试样进一步包括分别使所述第二和第三待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源分别对所述第二和第三待测试样中的粒子进行照射，以测得第二和第三待测试样的光散射信号和荧光信号；其中，所述光散射信号包括来自第二待测试样的第二侧向光散射强度信号、以及来自第三待测试样的第三侧向光散射强度信号；所述荧光信号包括来自第二待测试样中第一荧光染料的第二荧光强度信号和来自第三待测试样中第二荧光染料的第三荧光强度信号；
所述获取粒子信息进一步包括根据所述第二待测试样的所述第二侧向光散射强度信号和所述第二荧光强度信号获得所述第二子样本中的粒子信息，和根据所述第三待测试样的所述第三侧向光散射强度信号和所述第三荧光强度信号获得所述第三子样本中的粒子信息；其中所述第二子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息，所述第三子样本中的粒子信息包括淋巴细胞信息、嗜碱性粒细胞信息和有核红细胞信息；以及，
所述原始细胞检测进一步包括：
当所述原始细胞信息显示所述样本不含原始细胞时，根据所述第二子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息；或者
当所述原始细胞信息显示所述样本含有原始细胞时，根据所述第二子样本中的粒子信息，结合所述原始细胞信息和所述第三子样本中的粒子信息中的淋巴细胞信息，消除所述原始细胞信息对所述第二子样本中的粒子信息的影响，获得中性粒细胞、嗜酸性粒 细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息。
根据又一些实施方式，所述方法还包括对样本进行常规检测。
其中，根据一种实施方式，所述常规检测包括以下步骤：
吸取所述样本的至少部分作为第一初始子样本；
处理所述第一初始子样本，包括将所述第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样，其中所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同；
使所述第一初始待测试样中的粒子逐个通过光学检测装置的检测区并且利用所述光学检测装置的光源对所述第一初始待测试样中的粒子进行照射，以获取所述第一初始待测试样中粒子的光散射信号和荧光信号，所述光散射信号包括来自第一初始待测试样的第一初始侧向光散射强度信号；所述荧光信号包括来自第一初始待测试样中第一荧光染料的第一初始荧光信号；
根据所述第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括白细胞信息；和
当所述白细胞信息异常时，发出原始细胞报警，并可选地发出所述操作指令。
根据又一种实施方式，所述常规检测包括以下步骤：
分别吸取所述样本的至少部分作为第一初始子样本和第二初始子样本；
分别处理所述第一和第二初始子样本，包括将所述第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样，和将所述第二初始子样本与第五试剂和第六试剂混合以获得第二初始待测试样，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同，以及所述第二荧光染料不同于所述第一荧光染料；
分别使所述第一和第二初始待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源分别对所述第一和第二初始待测试样中的粒子进行照射，以获取所述第一和第二初始待测试样中粒子的光散射信号和荧光信号，所述光散射信号包括来自第一初始待测试样的第一初始侧向光散射强度信号和来自第二初始待测试样的第二初始侧向光散射强度信号；所述荧光信号包括来自第一初始待测试样中第一荧光染料的第一初始荧光信号和来自第二初始待测试样中第二荧光染料的第二初始荧光信号；
根据所述第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括初始白细胞信息，和根据所述第二初始侧向光散射强度信号和第二初始荧光强度信号获得所述第二初始待测试样的粒子信息，所述粒子信息至少包括初始淋巴信息；和
当所述初始白细胞信息异常时，发出原始细胞报警，并可选地发出所述操作指令。
根据该实施方式，对所述样本进行上述原始细胞检测中进一步包括：
结合所述原始细胞信息和所述初始淋巴细胞信息，消除所述原始细胞对所述初始白细胞信息的影响，并获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
根据一些实施方式，所述常规检测在所述原始检测之前进行，进一步地可以根据常规检测的结果发出所述操作指令。
所述操作指令包括用户输入的指令或当发出原始细胞报警时自动触发的指令。
根据一些实施方式，所述方法进一步包括：对原始细胞进行计数。
根据一些实施方式，所述方法进一步包括：当所述原始细胞信息显示所述样本含有 原始细胞时，报警所述样本中存在原始细胞，或报警所述样本中存在原始细胞并输出原始细胞计数值。
根据较优的实施方式，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，所述方法进一步包括：根据所述第一前向光散射强度信号和所述抗体荧光强度信号确定所述原始细胞为淋系原始细胞和/或髓系原始细胞。
根据另一较优的实施方式，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，所述方法进一步包括：根据所述第一前向光散射强度信号和所述抗体荧光强度信号报警所述样本中存在淋系原始细胞和/或髓系原始细胞。
根据另外一些实施方式，所述方法进一步包括：根据所述第一侧向光散射强度信号和抗体荧光强度信号进一步获得所述样本中的白细胞信息，所述白细胞信息包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的信息
较优地，本公开中所述第一溶血剂与所述样本的作用时间为30～80秒，优选为50～60秒。
本公开中所述第一溶血剂能够使红细胞完全破碎，同时白细胞维持完整的细胞形态。
本公开中，较优地，所述第一溶血剂为选自季铵盐类阳离子表面活性剂、烷醚乙氧类非离子表活剂、烷醇聚氧乙烯醚类表面活性剂、烷基糖苷、三萜皂苷、甾族皂苷和二甘醇中的至少一种。优选地，所述第一溶血剂包括溶解在缓冲液中的二甘醇和甲醛。
本公开中，较优地，所述抗体为CD45。
本公开利用血细胞分析仪的白细胞四分类检测通道(即，DIFF通道)仅通过增加一个荧光标记的抗体试剂能够准确地从白细胞中区分出原始细胞，并能够进行准确计数，从而提供了一种方法简单、成本较低的利用常规血细胞分析仪就能够检测原始细胞的方法和血细胞分析仪。
本公开第二个目的提供一种样本分析仪，通过样本分配装置从样本容器中吸取样本并将吸取的至少部分样本分配至第一反应装置和/或第二反应装置，通过第一反应装置将样本与抗体试剂、第一溶血试剂制备形成第一待测试样；通过第二反应装置将样本与第一类试剂制备形成第二待测试样，通过第一试样输送组件将第一待测试样从第一反应装置输送至光学检测组件进行原始细胞检测，通过第一试样输送组件将第二待测试样从第二反应装置输送至光学检测组件进行白细胞分类计数检测，从而实现了一个样本分析仪可同时用于检测原始细胞和血常规的效果。
为此，本公开的第三方面提供一种样本分析仪，包括样本分配装置、光学检测装置、第一反应装置、第二反应装置、第一试样输送组件和控制器；
所述样本分配装置用于从样本容器中吸取样本，并将吸取的至少部分样本分配至第一反应装置和/或所述第二反应装置；
所述第一反应装置至少用于为样本与抗体试剂、第一溶血试剂提供反应场所以制备形成第一待测试样，其中，所述抗体试剂中的抗体能够与样本中原始细胞的表面抗原相结合；
所述第二反应装置至少用于为样本与第一类试剂提供反应场所以制备形成第二待测试样；
所述第一试样输送组件用于将所述第一待测试样从所述第一反应装置输送至所述光学检测组件，且用于将所述第二待测试样从所述第二反应装置输送至所述光学检测组件；
所述光学检测组件包括流动室、光发射部件和光接收部件，所述流动室用于供所述第一待测试样或所述第二待测试样在鞘液的裹挟下通过，所述光发射部件用于朝向所述流动室中的第一待测试样或所述第二待测试样发射光线，所述光接收部件用于接收所述光发射部件发射的光线经所述第一待测试样产生的第一光学信号以及用于接收所述光 发射部件发射的光线经所述第而待测试样产生的第二光学信号；
所述控制器被配置为：根据所述光接收部件反馈的所述第一光学信号，解析得到所述第一待测试样中的第一粒子信息，所述第一粒子信息包括原始细胞信息；
根据所述光接收部件反馈的所述第二光学信号，解析得到所述第二待测试样中的第二粒子信息，所述第二粒子信息包括中性粒细胞信息、嗜酸性粒细胞信息、单核细胞信息和淋巴细胞信息。
根据一种实施方式，所述样本分析仪还包括外壳，所述光学检测装置、所述第一反应装置和所述第二反应装置都位于所述外壳内。
根据一种实施方式，所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的样本量小于或等于60uL；且/或，所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的样本量在50uL±10uL之间。
根据一种实施方式，所述控制器还被配置为：在单个原始细胞检测项目的检测过程中，控制样本与抗体试剂在所述第一反应装置的反应时间小于或等于90s，优选为小于或等于60s。
根据一撰具体实施方式，所述抗体试剂为CD45试剂。
根据一种实施方式，所述样本分配装置还用于从第一试剂容器中吸取所述抗体试剂，并将吸取的所述抗体试剂分配至所述第一反应装置；或者，所述样本分析仪还包括第一试剂分配装置，所述第一试剂分配装置用于从第一试剂容器中吸取所述抗体试剂，并将吸取的所述抗体试剂分配至所述第一反应装置。
根据一种实施方式，所述抗体试剂中包含经荧光标记、且能够与原始细胞表面抗原相结合的抗体，所述第一光学信号包括由所述光发射部件发射的光线照射至所述第一待测试样产生的第一侧向散射光信号和由所述光发射部件发射的光线照射至所述第一待测试样经所述荧光标记产生的第一荧光信号；或者，所述抗体试剂中包含未经荧光标记、但能够与原始细胞表面抗原相结合的抗体，所述第二反应池用于为样本与所述抗体试剂、第一荧光试剂提供反应场所以制备形成所述第一反应液，所述第一光学信号包括由所述光发射部件发射的光线照射至所述第一待测试样产生的第一侧向散射光信号和由所述光发射部件发射的光线照射至所述第一待测试样经所述第一荧光试剂产生的第一荧光信号。
根据一种实施方式，所述第一反应装置用于为所述第一反应液与所述第一溶血试剂、第二荧光试剂提供反应场所以制备形成所述第一待测试样；
所述第一光学信号还包括由所述光发射部件发射的光线照射至所述第一待测试样经所述第二荧光试剂产生的第二荧光信号；
所述控制器根据所述光接收部件反馈的所述第一光学信号，解析得到所述第一待测试样中的第一粒子信息：根据所述第一侧向散射光信号和所述第一荧光信号解析得到第一原始细胞信息和第一血影区域信息，根据所述第二荧光信号解析得到第二血影区域信息；
将所述第一原始细胞信息和所述第一血影区域信息，扣减所述第二血影区域信息，得到第一待测试样的原始细胞信息。
根据一种实施方式，所述第二反应装置包括第三反应池，且包括第四反应池和第五反应池中的至少一者，所述第三反应池用于为样本与所述第一类试剂提供反应场所以制备形成所述第二待测试样，所述第四反应池用于为样本与第二类试剂提供反应场所以制备形成第三待测试样，所述第五反应池用于为样本与第三类试剂提供反应场所以制备形成第四待测试样；
所述第一试样输送组件还用于将所述第三待测试样或所述第四待测试样从所述第 二反应装置输送至所述流动室；
所述流动室还用于供第三待测试样或所述第四待测试样在鞘液的裹挟下通过，所述光发射部件还用于朝向所述流动室中的所述第三待测试样或所述第四待测试样发射光线，所述光接收部件还用于接收所述光发射部件发射的光线经所述第三待测试样产生的第三光学信号、或经所述第四待测试样产生的第四光学信号；
所述控制器还被配置为根据所述光接收部件的反馈信息执行如下的至少一个解析动作：
根据所述光接收部件反馈的所述第三光学信号，解析得到所述第三待测试样中的第三粒子信息，所述第三粒子信息至少包括嗜碱性粒细胞信息；
根据所述光接收部件反馈的所述第四光学信号，解析得到所述第四待测试样中的第四粒子信息，所述第四粒子信息包括网织红细胞信息。
根据一种实施方式，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至所述第一反应装置，将吸取的部分样本分配至所述第二反应装置，其中，所述样本分配装置从所述样本容器一次性吸取的样本量大于或等于分配至所述第一反应装置的样本量与分配至所述第二反应装置的样本量之和；或者，
当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取样本，将吸取的部分样本分配至所述第二反应装置，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至所述第一反应装置，其中，所述样本分配装置从所述样本容器一次性吸取的样本量大于或等于分配至所述第一反应装置的样本量与分配至所述第二反应装置的样本量之和；或者，
当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取第一份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第一份样本分配至所述第一反应装置，从样本容器中吸取第二份样本，将吸取的至少部分第二份样本分配至所述第二反应装置；或者，
当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取第一份样本，将吸取的至少部分第一份样本分配至所述第二反应装置，从样本容器中吸取第二份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第二份样本分配至所述第一反应装置。
根据一种实施方式，当所述样本分配装置从所述样本容器一次性吸取的样本至少用于分别分配至所述第一反应装置和所述第二反应装置时，所述样本分配装置从所述样本容器一次性吸取的样本量小于或等于200uL。
根据一种实施方式，所述第二反应装置至少包括第三反应池，所述控制器还被配置为：在所述第一粒子信息的所述原始细胞信息中，扣减所述第二粒子信息中的异常淋巴细胞数据，得到原始细胞的测量数据；且/或，
所述第二反应装置至少包括第四反应池，所述控制器还被配置为：在所述第一粒子信息的所述原始细胞信息中，扣减所述第三粒子信息中的嗜碱性粒细胞数据，得到原始细胞的测量数据。
根据一种实施方式，所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的抗体试剂的量小于5uL；且/或，
所述控制器还被配置为：在单个原始细胞检测项目的抗体试剂分配过程中，控制分配至所述第一反应装置的抗体试剂的量在5uL±2uL之间。
根据一种实施方式，所述样本分析仪还包括血红蛋白检测装置，所述样本分配装置还用于将吸取的至少部分样本分配至所述血红蛋白检测装置，所述血红蛋白检测装置用于对至少由样本与第四类试剂制成的第五待测试样进行血红蛋白参数测定，所述控制器还被配置为：根据所述血红蛋白检测装置反馈的信息，解析得到所述第五待测试样中的血红蛋白信息；且/或，
所述样本分析仪还包括阻抗检测装置，所述阻抗检测装置用于对至少由样本与第五类试剂制成的第六待测试样进行测定，所述控制器还被配置为：根据所述阻抗检测装置反馈的信息，解析得到所述第六待测试样中的血小板信息和/或红细胞信息。
根据一种实施方式，所述样本分析仪对一个样本容器内样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果；且/或，
所述样本分析仪对所述样本分配装置从样本容器中一次吸取的样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果。
根据一种实施方式，所述样本分析仪还包括识别装置，所述识别装置用于对样本容器进行识别，所述控制器还被配置为：根据所述识别装置的反馈信息，至少获取样本容器内样本的待测检测项目信息，根据所述待测检测项目信息，控制所述样本分析仪对样本容器内的样本进行原始细胞检测和/或血常规检测；且/或，
所述样本分析仪还包括人机交互装置，所述人机交互装置配置有原始细胞检测模式和血常规检测模式，所述控制器还被配置为：根据操作人员在所述人机交互装置选择的检测模式，控制所述样本分析仪对样本容器内的样本进行原始细胞检测和/或血常规检测。
附图说明
附图中，相同的附图标记指代相同或相似的部件。
图1为利用流式细胞术的血细胞分析仪通过核酸荧光染色进行白细胞分析的原理示意图。
图2为利用常规血细胞分析仪的DIFF通道对血液样本检测获得的侧向光散射强度和核酸荧光强度二维散点图，其中A图为正常(健康)血液样本的二维散点图，B图为含有原始细胞的血液样本的二维散点图。
图3为利用血细胞分析仪对血液样本检测获得的侧向光散射强度和荧光强度二维散点图，其中A图为对正常血液样本进行常规白细胞检测获得的侧向光散射强度-核酸荧光强度(SSC/核酸FL)的二维散点图；B图为根据本公开方法利用荧光标记抗体检测含原始细胞的血液样本获得的侧向光散射强度-抗体荧光强度(SSC/抗体FL)的二维散点图，其中框出的区域为原始细胞出现的区域。
图4为利用本公开的血细胞分析仪的DIFF通道对血液样本检测获得的侧向光散射强度和核酸荧光强度二维散点图与侧向光散射强度和抗体荧光强度二维散点图的对比，其中A图示出的是在溶血条件下利用含核酸荧光染料试剂处理含有原始细胞的样本的侧向光散射强度-核酸荧光强度二维散点图中通常应出现单核细胞和淋巴细胞的区域，B图示出了在溶血条件下利用含有荧光标记的CD45试剂处理相同的样本的抗体荧光强度-侧向光散射强度的二维散点图，框出的区域是原始细胞出现的位置。
图5为根据本公开一种实施方式的原始细胞检测方法的流程示意图。
图6为根据本公开另一种实施方式的原始细胞检测方法的流程示意图。
图7为根据本公开又一种实施方式的原始细胞检测方法的流程示意图。
图8示出了WNB通道检测获得的第三荧光强度-侧向光散射强度(FL/SSC)的二 维散点图中各粒子群的分布示意图。
图9示出了在抗体荧光强度-侧向光散射强度(抗体/SSC)的二维散点图中分别来自髓系(A图)和淋系(B图)样本中原始细胞粒子群对应到抗体荧光强度-前向光散射强度(抗体/FSC)的二维散点图(C图)中的位置。
图10是本公开一个实施方式提供的血细胞分析仪的液路系统示意图。
图11是本公开该实施方式提供的光学通道反应池、采样部件、光学检测单元、稀释液提供装置与流体动力装置的液路连接示意图。
图12是本公开该实施方式提供的阻抗计数检测单元、血红蛋白检测单元、稀释液提供装置与流体动力装置的液路连接示意图。
图13是根据实施例1检测获得的含有高含量未成熟粒细胞(IG)但不含原始细胞的样本的FL/SSC散点图。
图14是根据实施例2检测获得的含有原始细胞的样本的FL/SSC散点图。
图15是根据实施例2检测获得的含有原始细胞的样本的WNB通道FL/SSC散点图(A)和DIFF通道FL/SSC散点图(B)。
图16是根据实施例4检测获得的含有原始细胞的样本的CD45FL/SSC散点图。
图17是本公开另一实施方式提供的样本分析仪的组成示意图。
图18是本公开该实施方式提供的样本针的工作过程示意图。
具体实施方式
下面将结合本公开具体实施方式、实施例及附图，对本公开的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是本公开的一部分实施方式，而不是全部的实施方式。基于本公开中的实施方式，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施方式，都属于本公开保护的范围。
在整个说明书中，除非另有特别说明，本文使用的术语应理解为如本领域中通常所使用的含义。因此，除非另有定义，本文使用的所有技术和科学术语具有与本公开所属领域技术人员的一般理解相同的含义。若存在矛盾，本说明书优先。
本文中所有方向性指示(诸如上、下、左、右、前、后……)仅用于解释在某一特定姿态下各部件之间的相对位置关系、运动情况等，如果该特定姿态发生改变时，则该方向性指示也相应地随之改变。
在本文中涉及“第一”、“第二”等的描述仅用于描述目的，而不能理解为指示或暗示其相对重要性或者隐含指明所指示的技术特征的数量。由此，限定有“第一”、“第二”的特征可以明示或者隐含地包括至少一个该特征。另外，各个实施例之间的技术方案可以相互结合，但是必须是以本领域普通技术人员能够实现为基础，当技术方案的结合出现相互矛盾或无法实现时应当认为这种技术方案的结合不存在，也不在本公开要求的保护范围之内。
本文中提及的‘样本’，除非另外指明，是指血液样本、骨髓样本或体液样本。血液样本可以是外周血或静脉血。体液样本诸如为脑脊液、胸水、腹水、心肌液、滑液、腹膜透析液或腹膜清洗得到的液体等。
本文中提及的“原始细胞”或在本文中表示为“BLAST”，二者可替换使用，均指各种血细胞开始发育阶段的细胞。在健康人的血液中不会出现原始细胞，但是患有一些疾病，特别是血液疾病的人的血液中会含有原始细胞。
本文中提及的‘溶血剂’，除非另外指明，是指能够使红细胞裂解，同时使白细胞保持基本细胞形态而细胞膜受到一定程度的破坏的试剂。例如，可用于本公开的溶血剂可以选自表面活性剂、烷基糖苷、三萜皂苷、甾族皂苷等。
本文中提及的“抗体”除非另外指明，是指能够与原始细胞的细胞膜表面抗原结合的 抗体，特别是单克隆抗体。所述抗体与原始细胞的细胞膜表面抗原可以是非特异性结合，也就是说所述抗体还能够与血液中其他细胞，如白细胞的细胞膜表面抗原结合。但是所述抗体与不同血液细胞表面抗原结合的情况(如牢固度、数量、部位等)不同，因而可以由此获得不同的信号。
本文中提及的‘荧光标记的抗体’指结合有可用于检测的荧光标记物的上述抗体。所述荧光标记物可以根据不同的检测平台选择具有合适的激发波长的那些荧光物质。
本文中提及的‘荧光染料’，除非另外指明，是指能够特异性与细胞内物质(如DNA、RNA或蛋白质)结合并在激发光的激发下发出特定波长荧光的染料。
通常，血液中白细胞的检测可使用全自动血液细胞分析仪。具体地可以通过鞘流阻抗法、激光散射法、结合核酸荧光染色的流式细胞技术等对样本中的白细胞进行分类和计数。其中，利用结合核酸荧光染色的流式细胞技术(DIFF通道)的检测，是在溶血条件下利用核酸荧光染色，进而通过光信号进行分析，其原理如图1所示。血细胞分析仪吸取血液样本后，样本首先经溶血剂和核酸荧光染料处理，红细胞被溶血剂破坏破碎，白细胞仍维持细胞形态，核酸荧光染料可进入白细胞内，与细胞内的核酸特异性结合。接着，利用流式细胞术，样本中的细胞逐个通过激光检测孔，由光源发出的激光束照射到正在通过检测孔的细胞时，产生各个角度的散射光，同时与核酸结合的荧光染料被激光激发，发出特定波长的荧光。根据细胞本身的特性(如体积、染色程度、细胞内容物大小及含量、细胞核密度等)所产生的不同角度的散射光和荧光的强度不同，这些散射光和荧光信号经光学检测器接收后可以获得细胞结构和组成的相关信息。其中前向光散射强度(Forward scatter，FS)反映细胞的体积，侧向光散射强度(Side scatter，SS)反映细胞内部结构的复杂程度，荧光强度(Fluorescence，FL)反映细胞中核酸物质(包括DNA和RNA)的含量。利用这些参数可以对样本中的白细胞进行分类和计数。通常在DIFF通道利用侧向光散射强度和荧光强度的二维信号数据形成二维散点图，可以获得淋巴细胞、单核细胞、嗜酸性粒细胞和中性粒细胞的四分类计数(参见图2中的A图)。当患者外周血中存在原始细胞时，原始细胞也同时被荧光染料染色，而其粒子信息在SS-FL二维散点图中往往会覆盖淋巴细胞与单核细胞区域(称为“原始细胞敏感区域”，参见图2中的B图)，造成淋巴细胞、单核细胞、原始细胞三者无法区分，进而淋巴细胞和单核细胞无法准确定量。而且当前商品化的血细胞分析仪只能提供原始细胞报警提示而无法进行原始细胞的区分和准确计数，而且报警提示的敏感性和特异性不高，假阳性和假阴性的情况均可能存在。
本发明人发现，与上述传统的白细胞检测中利用核酸荧光染料不同，利用能够与原始细胞表面抗原结合的单克隆抗体可以通过单抗与不同细胞表面抗原的结合程度不同，同时结合反映细胞内部结构复杂度的侧向散射光，能够直接使原始细胞区分于其他白细胞。参考图3，该图为利用血细胞分析仪，对经过溶血剂和含有核酸荧光染料处理过的正常血液样本的检测结果(A图)和对经过溶血剂和含有荧光标记的抗体处理过的含有原始细胞的血液样本的检测结果(B图)。其中参见B图，在侧向光散射强度-抗体荧光强度的二维散点图中，不规则图形框出的区域为原始细胞出现的区域(BLAST)。从B图能够看出通过抗体荧光信号能够使原始细胞完全区分于其他白细胞，从而能够实现对原始细胞的准确定位。
进一步参考图4，其中对比了当样本中存在原始细胞时，常规的核酸荧光法进行白细胞分析中获得的二维散点图和利用本公开方法分析获得的二维散点图。B图示出了在溶血条件下利用含有荧光标记的抗体试剂处理含有原始细胞的样本后的抗体荧光强度-侧向光散射强度的二维散点图，框出的区域是原始细胞出现的位置。而A图示出的是在溶血条件下利用含核酸荧光染料试剂处理相同的样本后通常应能够区分单核细胞和淋巴细胞的区域，完全无法区分原始细胞、单核细胞和淋巴细胞。
由此，本发明人提出了一种能够将原始细胞与其他白细胞区分开，并能够对原始细胞进行准确计数的血细胞分析仪以及原始细胞检测方法。
根据本公开的实施方式，提供了一种样本分析方法。
所述方法包括接收操作指令，并对样本进行原始细胞检测。操作指令可以是用户选择的指令(例如对于已知可能含有原始细胞的样本，可直接选择进行原始细胞检测；或者仪器发出原始细胞报警后，也可选择是否进行原始细胞检测)，也可以是血细胞分析仪检测中发出原始细胞报警直接触发的。
对样本进行原始细胞检测，参见图5，可包括以下步骤。
S10，吸取样本，包括吸取所述样本的至少部分作为第一子样本。
S20，处理样本，包括将所述第一子样本与第一类试剂混合以获得第一待测试样；其中所述试剂包括第一试剂和第二试剂，所述第一试剂包括第一溶血剂，所述第一溶血剂与所述第一子样本的混合时间不超过2分钟，所述第二试剂包括荧光标记的抗体，所述抗体能够与原始细胞的表面抗原相结合。
S30，检测待测试样，包括使所述第一待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源对所述第一待测试样中的粒子进行照射，以测得第一待测试样的光散射信号和荧光信号，所述光散射信号包括来自第一待测试样的第一侧向光散射强度信号，所述荧光信号包括来自所述经荧光标记的抗体的抗体荧光强度信号。
S40，获取粒子信息，包括根据所述第一侧向光散射强度信号和所述抗体荧光强度信号获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括原始细胞信息。该方法可以利用以下详述的血细胞分析仪的DIFF通道，也可以增加新的单独的检测通道。
在该实施方式中，S10吸取样本的步骤，可通过吸样装置定量吸取部分样本，作为第一子样本以用于后续的原始细胞检测。该吸取样本的步骤还可包括多次吸取部分样本获得多个子样本，用于不同的检测，如白细胞分析、血小板分析、血红蛋白分析等等。该步骤也可以包括将一次吸取的样本分为若干子样本用于不同的检测。
在S20处理样本的步骤中，第一子样本与第一和第二试剂混合，进行溶血和抗体-抗原反应，以制备第一待测试样。第一和第二试剂可以任何顺序先后与样本混合，也可以同时与样本混合。将混合后的样本孵育一定时间，使红细胞破碎，并使荧光标记的抗体与细胞表面的抗原结合。
该步骤的含第一溶血剂的第一试剂可对红细胞进行快速溶血，例如可在约2分钟内完成溶血，通常溶血时间在1分钟以内，更通常在30～80秒，甚至50～60秒。第二试剂中荧光标记的抗体与细胞表面的抗原相结合的反应通常需要在约42～45℃温度下进行约45～60秒时间。根据选择的抗体不同，以及孵育温度不同，反应时间会有所不同。
第一试剂中的第一溶血剂可以使用与常规白细胞检测中DIFF通道使用的那些溶血剂也可以使用不同于DIFF通道使用的溶血剂。第一溶血剂可以一种溶血剂或也可以是多种溶血剂的组合。根据一种实施例，第一溶血剂可以包括选自季铵盐类阳离子表面活性剂、烷醚乙氧类非离子表活剂、烷醇聚氧乙烯醚类表面活性剂、烷基糖苷、三萜皂苷、甾族皂苷和二甘醇中的至少一种具体实例可为具有通式I的糖苷类化合物：
R-(CH2)n-CH3   (I)
其中，R选自由单糖、去氧单糖和多糖所组成的组，n为5～17的整数。中国专利申请CN111602052A中公开的溶血剂均可作为本公开的第一溶血剂。该专利文献的全部内容通过引用并入本文。根据另一些实施例，第一溶血剂可以包括二甘醇，优选进一步包括对细胞起固定作用的甲醛。
根据另一些实施例，第一溶血剂可包括阳离子表面活性剂、非离子表面活性剂、以 及芳香族有机酸和/或其盐。其中，芳香族有机酸可选自由对苯二甲酸、邻苯二甲酸、羟基苯甲酸、乙酰水杨酸、对氨基苯甲酸、苯磺酸、对甲苯磺酸和羟基苯磺酸所组成的组中的一种或多种。阳离子表面活性剂选自季铵盐和吡啶鎓盐中的至少一种。非离子表面活性剂选自聚氧乙烯类表面活性剂、斯潘和吐温中的至少一种。
所述第一试剂中还可含有缓冲剂(如磷酸及其盐、柠檬酸及其盐、乙酸及其盐等)、防腐剂(如叠氮钠、ProClin系列等)、金属螯合剂(如EDTA的钠盐)、渗透压调节剂(如氯化钠等)等组分。
可用的第一试剂可以是已商品化的用于白细胞检测的溶血剂，例如M-68P LD溶血剂。也可以是其他溶血剂，例如溶解在缓冲溶液中的二甘醇和甲醛。
第二试剂中荧光标记的抗体可以是能够与原始细胞表面抗原结合的那些连接了合适的荧光标记物的抗体，特别是单克隆抗体。
本公开对抗体没有特别限制，只要与各类细胞结合力不同，能够通过抗体荧光和侧向光散射强度将原始细胞区分于其他白细胞的均可使用，例如可以是CD45、CD71、CD34中的一种，也可以是两种或更多种的组合。优选仅使用一种抗体，更优选抗体为CD45。
本公开对所述荧光标记物也没有特别限制，只要与抗体结合后不影响抗体与抗原结合，也不会影响其他试剂(如溶血剂)使用即可。可根据血细胞分析仪所使用激光光源的波长来选择能够被该光源激发的那些荧光物质作为标记物。这些荧光标记物是本领域技术人员所熟知的。可列举的荧光标记物包括FITC(蓝光激发)、APC(红光激发)，但不限于此。可使用已经商品化的荧光标记的抗体，例如：CD45-FITC、CD45-APC等。
第二试剂中荧光标记的抗体的浓度可为100～200μg/mL，溶剂通常是pH为7.2的磷酸盐缓冲液。第二试剂中还可包括叠氮化钠、牛血清白蛋白。
第一试剂与样本的体积混合比例可为常规溶血处理所使用的量，如50:1。例如可用约1mL的第一试剂与约20μL的样本混合。第二试剂与样本的体积混合比例可为1:20。
在S30检测待测试样的步骤中，已经完成红细胞溶血和抗体-抗原的反应的第一待测试样输送到光学检测单元进行光学检测。在光学检测单元，每个粒子逐一通过流动室的检测区，在激光光源的照射下，产生不同方向的散射光，同时由于细胞表面结合了抗体，抗体携带的荧光标记物被激发，发出荧光。每个粒子的光散射信号以及荧光信号被布置在检测区周围的信号收集装置收集，并传输到处理器。在该实施方式中第一待测试样的光散射信号至少包括第一侧向光散射强度信号。在其他实施方式中，同时还可收集第一前向光散射强度信号，甚至进一步根据需要收集其他角度的光散射强度信号。在该实施方式中第一待测试样的荧光信号为抗体携带的荧光标记物发出的抗体荧光强度信号。
根据本公开的一些实施方式，对原始细胞的检测可利用常规白细胞检测的DIFF通道，仅需将用于白细胞检测的荧光染色试剂替换为抗体荧光试剂。根据其他实施方式，对原始细胞的检测可利用单独的抗体通道进行检测，而白细胞仍使用DIFF通道进行检测。
在S40获取粒子信息的步骤中，根据接收的第一侧向光散射强度信号和荧光强度信号可形成SSC-抗体FL的二维散点图，并获得包括原始细胞信息的第一子样本的粒子信息。仍参考图3和图4的B图，可以看到在包含抗体荧光信号的坐标中，原始细胞的粒子团能够显著区别于其他白细胞粒子团，从而能够获取准确的原始细胞信息。对于来自健康受试者的样本，在原始细胞粒子团区域可能检测不到任何粒子(即，原始细胞百分比为0％)或粒子数量极少。对于来自患者的样本(如图4中B图所示)，则可进一步获得准确的原始细胞分类和计数。
根据本公开，所述方法在获取原始细胞的分类计数的同时还可消除原始细胞对白细胞检测的影响进一步获取白细胞准确计数，具体可通过多种实施方式实现。
根据第一种实施方式，可利用血细胞分析仪的DIFF通道一次制样、一次检测以同时获得原始细胞和白细胞信息，并能够获得准确的原始细胞计数，而且获得白细胞四分类计数而不会受到样品中原始细胞的影响。该实施方式尤其适用于装配了双荧光检测器的血细胞分析仪。本领域技术人员应理解，在该方法中，标记抗体的荧光标记物发射的荧光波长应当不同于白细胞计数中使用的荧光试剂发射的荧光波长。具体地，参考图6，所述原始细胞的检测方法可包括以下步骤。
S110，吸取部分样本，作为第一子样本。该步骤与上述实施方式中的S10步骤相同。
S120，处理样本，将第一子样本与包括第一溶血剂的第一试剂、包括荧光标记的抗体的第二试剂和包括第一荧光染料的第三试剂混合，获得第一待测试样。该步骤与上述实施方式中的S20步骤的区别在于第一子样本还与包括第一荧光染料的第三试剂混合。在该步骤中，第一、第二和第三试剂可以以任何顺序依次与第一子样本混合，也可以同时混合。
第一和第二试剂如上所述，与样本的混合方法也如上所述。
第三试剂可以是常规用于DIFF通道白细胞检测所使用的核酸荧光染料试剂。对于第三试剂中的第一荧光染料没有特别限制，可根据所使用的血细胞分析仪光源的不同选择具有合适激发波长的荧光染料，该第一荧光染料为核酸荧光染料。商品化的核酸荧光染料及一些专利申请中已经公开的核酸特异性荧光染料均可用于所述方法。
其中商品化的核酸荧光染料，可列举的有Thermofisher公司的SYTO系列核酸染料。此外，中国专利申请No.：CN201010022414.6中公开的荧光染料、No.：CN200910109215.6中公开的花青素类染料、CN200810216864.1中公开的荧光染料等，均可用于本公开。以上专利文献的全部内容通过引用并入本文。
可将第一荧光染料溶解在适宜的有机溶剂(如乙醇、甲醇、乙二醇等)中形成第三试剂。第三试剂可使用已商业化的那些，如M-68P FD染色液。本公开对第三试剂没有特别限制。
第三试剂与样本的体积混合比例可为常规利用DIFF通道进行白细胞检测时的混合比例，如可为1:1。
经步骤S120处理后获得的第一子样本中红细胞已破碎，仍保持完整细胞形态的血细胞，如白细胞，以及可能存在的原始细胞，胞内核酸物质已被染色，同时细胞膜上也结合了荧光标记的抗体。
S130，检测待测试样，与上述实施方式相同，第一待测试样中的粒子逐个通过流动室的检测区，在激光光源的照射下，产生不同方向的散射光，同时由于细胞表面结合了抗体，抗体携带的荧光标记物被激发，发出抗体荧光，不同之处在于细胞内部的核酸结合了第一荧光染料，该染料具有与核酸结合后荧光量子产率显著增加的特点，因而也被激发，发出荧光。在该实施方式中第一待测试样的光散射信号至少包括第一侧向光散射强度信号。在其他实施方式中，同时还可收集第一前向光散射强度信号，甚至进一步根据需要收集其他角度的光散射强度信号。在该实施方式中第一待测试样的荧光信号为抗体携带的荧光标记物发出的抗体荧光强度信号以及与核酸结合的第一荧光染料发出的第一荧光强度信号。
S140，获取粒子信息，包括根据第一侧向光散射强度信号和抗体荧光强度信号获得的原始细胞信息；和根据第一侧向光散射强度信号和第一荧光强度信号获得白细胞信息。
在该实施方式中，由于细胞同时结合了荧光标记的抗体和第一荧光染料，因而每个粒子可被测得至少四个维度的信号，能够利用不同维度的信号获得更多粒子的信息。如前所述，利用第一侧向光散射强度信号和抗体荧光强度信号能够直接获得的原始细胞的准确分类和计数。但是利用第一侧向光散射强度信号和第一荧光强度信号获得的白细胞 信息会受到样本中可能存在的原始细胞影响，导致淋巴细胞、单核细胞和原始细胞难以区分(参见图4的A图)。
为获得准确的白细胞四分类信息，该步骤中需要利用原始细胞信息。当检测到原始细胞占比为0％，即，样本中不含原始细胞，可直接根据第一侧向光散射强度信号和第一荧光强度信号获得的中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数(S141)。而当检测到原始细胞占比大于0％时，即，样本中含有原始细胞，则从所述第一子样本的粒子信息中扣除所述原始细胞的影响(即，利用第一侧向光散射强度信号和抗体荧光强度信号划分出原始细胞粒子的信息后，在第一侧向光散射强度信号和第一荧光强度信号的散点图中扣除原始细胞信息)，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数(S142)。
此外，在该S141和S142步骤中，还可进一步获得未成熟粒细胞信息和/或异型淋巴细胞信息，如果样本中存在这些细胞的话。
根据第二种实施方式，分别制备三个待测试样，一个用于原始细胞检测，一个用于DIFF通道的四分类白细胞检测，还有一个用于WNB通道的有核红细胞、嗜碱性粒细胞和淋巴细胞等的检测。该实施方式尤其适用于仅装配了一个荧光检测器的血细胞分析仪。具体地，参考图7，所述原始细胞的检测方法可包括以下步骤。
S210，吸取样本作为第一、第二和第三子样本。该步骤中，可一次吸样分为三个血段，分别检测；也可以分若干次吸样。
S220，分别处理各子样本。该步骤中对三个子样本分别进行处理。
其中，S221，将第一子样本与上述第一试剂和第二试剂混合获得第一待测试样。该步骤与上述实施方式中的S20步骤相同。
S222，将第二子样本与上述第三试剂，以及第四试剂混合以获得第二待测试样。该步骤与常规利用DIFF通道进行白细胞检测中的样本处理步骤相同。
S223，将第三子样本与第五试剂和第六试剂混合以获得第三待测试样。该步骤与常规利用WNB通道进行有核红细胞、嗜碱性粒细胞和淋巴细胞检测中的样本处理步骤相同。
其中第四试剂包括第二溶血剂，第五试剂包括第二荧光染料，第六试剂包括第三溶血剂。如前所述，所述第二溶血剂与所述第一溶血剂可以相同也可以不同，所述第三溶血剂与第一和第二溶血剂均不同。所述第二荧光染料不同于所述第一荧光染料。其中第五和第六试剂是常规用于WNB通道检测的荧光试剂和溶血剂。
第四试剂包括第二溶血剂。所述第二溶血剂是常规白细胞检测中DIFF通道使用的那些溶血剂。例如，第一溶血剂可以一种溶血剂或也可以是多种溶血剂的组合。根据一种实施例，第一溶血剂可以表面活性剂，例如阳离子表面活性剂(如季铵盐类阳离子表面活性剂)、非离子表面活性剂(例如烷醇聚氧乙烯醚类表面活性剂)。在另一些实施例中，第一溶血剂可以是烷基糖苷类、三萜皂苷类或甾族皂苷类物质。具体实例可为具有通式I的糖苷类化合物：
R-(CH2)n-CH3   (I)
其中，R选自由单糖、去氧单糖和多糖所组成的组，n为5～17的整数。中国专利申请CN111602052A中公开的溶血剂均可作为本公开的第一溶血剂。该专利文献的全部内容通过引用并入本文。
与前述第一试剂类似，所述第四试剂中还可含有缓冲剂、防腐剂、金属螯合剂、渗透压调节剂等组分。
可用的第一试剂可以是已商品化的用于白细胞检测的溶血剂，例如M-68P LD溶血剂。
第四试剂与样本的体积混合比例是常规DIFF通道检测中使用的比例，如可为50:1。
第五试剂包括第二荧光染料。所述第二荧光染料是能够与蛋白结合的一类染料。所述第二荧光染料的实例可为具有下式结构的化合物：
其中可以各自独立为
R7、R8和R9可选自H、卤素、氰基、羟基、C1-18烷基、C1-18烷基磺酸基、磺酸基和C1-15烷基COOR10，其中R10为H或C1-6的烷基；
R5、R6各自独立选自C1-18烷基COOR11、C1-18烷基OR11和苄基，其中苄基可以被选自卤素、羟基、巯基、氰基、硝基、烷基、芳基、烷氧基、杂环基、卤代烷基、氨基、烷基氨基、酰胺基和羧基中的基团所取代，并且R5和R6不同时为苄基；
R11每次出现时可以独立地为H、C1-18烷基或者苯基，其中苯基可以被选自卤素、羟基、巯基、氰基、硝基、烷基、芳基、烷氧基、杂环基、卤代烷基、氨基、烷基氨基、酰胺基和羧基的基团所取代；
X为-CH2-、-C(CH3)2-、-O-、-S-或-Se-；
Z-为阴离子。
第二荧光染料可以例如50μg/mL浓度溶解在合适的溶剂(如甲醇、乙醇、乙二醇等)中作为第五试剂。用作第二荧光染料的实例可以是已经商业化的那些，如M-68P FN染色液。
第六试剂包括第三溶血剂。所述第三溶血剂与第一和第二溶血剂不同。第三溶血剂可包括选自阳离子表面活性剂和非离子表面活性剂中的至少一种表面活性剂。
所述阳离子表面活性剂可选自具有式1所示结构的季铵盐型阳离子表面活性剂，具体实施例结构式为：
其中R1为C原子数6-14的烷基或链烯基；
R2、R3为C原子数1-4的烷基或链烯基；
R4为C原子数1-4的烷基或链烯基或苄基；
B为卤素原子。
上述阳离子表面活性剂优选十二烷基三甲基氯化铵、十六烷基三甲基季铵溴化铵、十八烷基二甲基苄基季铵氯化铵。
所述非离子表面活性剂可选自辛基苯基聚氧乙烯醚、聚氧乙烯十六烷基醚。
第三溶血剂也可包括芳香族有机酸或其盐，如水杨酸及其盐，苯甲酸及其盐。
作为第六试剂的第三溶血剂可以为商品化的用于白细胞检测的溶血剂,例如(M-68P LN溶血剂)，第三溶血剂与第一或第二溶血剂相比可为酸性溶血剂(pH3.0)。第六试剂与第一试剂类似，还可以包括缓冲剂、防腐剂、渗透压调节剂、络合剂等。
第五和第六试剂与样本的体积混合比可按照常规WNB通道处理样本的方式。如第五试剂与样本的混合比例可为1:1，第六试剂与样本的混合比例可为50:1。
对子样本的处理可以在同一混合室中依次进行，也可以在不同的混合室中进行。
S230，分别检测各待测试样。该步骤中对三个待测试样分别进行检测。
其中，S231，对第一待测试样进行检测。该步骤与前述实施方式中的S30步骤相同。
S232，对第二待测试样进行检测，获得包括第二侧向光散射强度信号的散射光信号，以及包括来自第一荧光染料的第二荧光强度信号。该步骤与常规利用DIFF通道对样本进行检测的步骤相同。
S233，对第三待测试样进行检测，获得包括第三侧向光散射强度信号的散射光信号，以及包括来自第二荧光染料的第三荧光强度信号。该步骤与常规利用WNB通道对样本进行检测的步骤相同。
在该检测步骤中，三个待测试样依次通过光学检测单元进行检测，从而获得各待测试样中粒子的光学信号。这些光学信号被传输到处理器，以进行下一步骤。
接着，在S240中，获取各待测试样的粒子信息。
S241，针对第一待测试样，与前述实施方式中S40步骤相同，根据第一侧向光散射强度信号和抗体荧光强度信号获得原始细胞信息。
S243，针对第三待测试样，根据第三侧向光散射强度信号和第三荧光强度信号，获得至少包括淋巴细胞、嗜碱性粒细胞和有核红细胞的粒子信息。该步骤与常规使用WNB通道进行样本分析中的检测步骤相同。
在WNB通道，可以获得嗜碱性粒细胞和有核红细胞的分类计数，此外还能获得准确的淋巴细胞计数。由于所用的第二荧光染料的特点，样本中即便存在原始细胞，也不会干扰淋巴细胞的分类计数。参见图8，其中示出了WNB通道检测获得的荧光强度-侧向光散射强度的二维散点图中各粒子群的分布示意图。从图8可见，原始细胞出现的位置与异常淋巴细胞重合在一起，但是与正常淋巴细胞能够较好地区分开。因此，在WNB通道可以获得针对第三待测试样的准确的淋巴细胞百分比。此外，嗜碱性粒细胞和有核红细胞的粒子群也不受原始细胞的影响，因此还能获得准确的嗜碱性粒细胞和有核红细胞的分类计数。
S242，针对第二待测试样，根据第二侧向光散射强度信号和第二荧光强度信号获得至少包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的粒子信息。该步骤中，与前述第一实施方式类似，四分类白细胞中淋巴细胞和单核细胞的分类可能受到原始细胞的干扰，而无法进行分类计数。
在该实施方式中，针对原始细胞信息的不同情况进一步包括以下步骤。
S2421，当检测到原始细胞占比为0％，即样本中不含原始细胞，可直接根据第一侧向光散射强度信号和第一荧光强度信号获得的中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数。
S2422，当检测到原始细胞占比大于0％时，即样本中含有原始细胞，则结合来自第一待测试样获得的原始细胞信息和来自第三待测试样获得的淋巴细胞信息，消除所述原始细胞信息对所述第二子样本中的粒子信息的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数。
此外，在该S2421和S2422步骤中，还可进一步获得未成熟粒细胞信息和/或异型淋巴细胞信息，如果样本中存在这些细胞的话。
根据本公开还能提供样本中原始细胞是髓系原始细胞还是淋系原始细胞的信息。参见图9，其中示出了在抗体荧光强度-侧向光散射强度(抗体/SSC)的二维散点图中分别来自髓系和淋系样本中原始细胞粒子群对应到抗体荧光强度-前向光散射强度(抗体/FSC)的二维散点图中的位置。从图9可以清晰地看到在抗体/FSC的二维散点图中来自髓系样本中的原始细胞和来自淋系样本中的原始细胞有明确的位置关系。因此，可以进一步根据抗体荧光强度信号和前向光散射强度信号确定原始细胞为髓系和/或淋系原始细胞。
在上述各实施方式中，均可进一步利用第一待测试样的粒子第一前向光散射嵌段信号和荧光强度信号来确定被测样本中原始细胞为髓系和/或淋系原始细胞；或者报警被测样本中存在淋系原始细胞和/或髓系原始细胞。
根据本公开，在上述各实施方式中，可直接对被测样本中原始细胞进行计数而不需判断是否存在原始细胞。或者，当检测到被测样本中存在原始细胞时，报警被测样本中存在原始细胞，或报警被测样本中存在原始细胞并输出原始细胞计数值。
进一步的，本发明人还发现，在本公开的利用荧光标记的抗体进行检测的方法中，在侧向光散射强度-抗体荧光强度二维坐标的散点图中，可以很好地区分包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的细胞分类信息(参见图3的B图)。
基于此，在上述各实施方式中，本公开的原始细胞检测方法还包括针对第一待测试样获得的第一侧向光散射强度信号和抗体荧光强度信号获得样本中的白细胞分类信息，所述白细胞信息包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的信息。进一步地，所述方法包括对淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的分类计数。
根据本公开的又一种实施方式，所述样本分析方法还包括对样本进行常规检测。
所述常规检测包括对待测样本中的各种细胞进行分析并分类计数，还包括对一些生化指标的分析。根据一些实施方式，常规检测可以是临床中的血常规检测。根据具体的实施方式，所述常规检测可以包括对红细胞、白细胞、血小板、血红蛋白等项目的检测和分析。
如果该常规检测发现样本可能存在原始细胞，则可选择启动对原始细胞的检测；如果该常规检测未发现样本可能存在原始细胞，则不必启动对原始细胞的检测。该实施方式适用于未确定患病或未存疑患病的受试者，例如大量的普通门诊患者或进行健康体检的患者，以节约检测的时间和成本。
根据一些实施方式，本公开的样本分析方法包括对样本进行利用DIFF通道的白细胞检测。在该实施方式中，所述常规检测包括以下步骤。
首先，吸取样本的至少部分作为第一初始子样本。
接着，处理第一初始子样本。该步骤包括将所述第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样。所述第三试剂和第四试剂如上所述。
然后，使第一初始待测试样中的粒子逐个通过光学检测单元的检测区并且利用所述光学检测单元的光源对第一初始待测试样中的粒子进行照射，以获取第一初始待测试样中粒子的光散射信号和荧光信号，所述光散射信号包括来自第一初始待测试样的第一初始侧向光散射强度信号；所述荧光信号包括来自所述第一荧光染料的第一初始荧光信号。
最后，根据第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括白细胞信息；和当所述白细胞信息异常时，发出原始细胞报警。在该步骤中，如前所述，如果样本中存在原始细胞，则在荧光强度-侧向光散射强度的二维散点图中，淋巴细胞和单核细胞的粒子群与原始细胞粒子群重叠，导致淋巴细胞和单核细胞无法分类。在常规的血细胞分析仪的检测中，将针对该情况发出样本中可能存在原始细胞的报警。
进一步地，基于报警，可以由用户选择发出操作指令，或者由报警直接触发操作指令，根据上述各实施方式中任意一种方法的进行原始细胞的检测。
根据另一些实施方式，本公开的样本分析方法包括对样本进行利用DIFF通道的四分类白细胞检测以及利用WNB通道的有核红细胞、嗜碱性粒细胞和淋巴细胞检测。在该实施方式中，所述常规检测包括以下步骤。
首先，分别吸取样本的至少部分作为第一初始子样本和第二初始子样本。
其次，分别处理第一和第二初始子样本，包括将第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样，和将第二初始子样本与第五试剂和第六试剂混合以获得第二初始待测试样。其中，所述第三、第四、第五和第六试剂分别如上所述。
然后，分别使第一和第二初始待测试样中的粒子逐个通过光学检测单元的检测区，并且利用所述光学检测单元的光源分别对所述第一和第二初始待测试样中的粒子进行照射，以获取所述第一和第二初始待测试样中粒子的光散射信号和荧光信号，所述光散射信号包括来自第一初始待测试样的第一初始侧向光散射强度信号和来自第二初始待测试样的第二初始侧向光散射强度信号；所述荧光信号包括来自第一初始待测试样中第一荧光染料的第一初始荧光信号和来自第二初始待测试样中第二荧光染料的第二初始荧光信号。
最后，根据所述第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括初始白细胞信息，和根据所述第二初始侧向光散射强度信号和第二初始荧光强度信号获得所述第二初始待测试样的粒子信息，所述粒子信息至少包括初始淋巴信息；和当所述初始白细胞信息异常时，发出原始细胞报警。
基于报警，可选择发出操作指令进行原始细胞的检测。在该实施方式中，可进行上述任何一种实施方式的原始细胞检测。
此外，在该实施方式中，还可仅进行如前述图5所示实施方式的原始细胞检测。在对原始细胞的检测结束后，结合原始细胞检测中获得的原始细胞信息和常规检测中获得的初始淋巴细胞信息，消除原始细胞对常规检测中获得的初始白细胞信息的影响，并获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数。进一步的，如果样本中存在的话还可获得未成熟粒细胞信息和/或异型淋巴细胞信息。
本公开进一步提供一种血细胞分析仪。
根据一种实施方式，提供血液分析仪，包括吸样装置、试剂供应装置、混合室、光源、光学流动室、光学检测器、处理器和编程有计算机应用程序的非暂时性计算机可读存储介质。
吸样装置用于吸取样本，并将所吸取的样本输送到所述混合室。吸取样本可以根据需要吸取部分样本用于上述样本分析方法的各实施方式中的某一种或某几种测试。对于多种测试，吸样装置可以一次性吸取一定量的样本，分为若干子样本，分别进行样本的处理和检测；也可以多次吸取一定量的样本，分别作为各子样本，并分别进行处理和检测。
试剂供应装置用于提供试剂并输送到混合室。试剂供应装置可以有多个，以分别用于提供不同试剂。如上样本分析方法的各实施方式，试剂供应装置用于根据不同的检测需要提供进行样本处理所需的，例如第一至第六试剂中的一种或多种。
混合室用于样本与试剂的混合和反应。混合室也可以有多个，可进行不同检测所需的各子样本的处理。此外，同一混合室也可先后进行不同子样本的处理。
混合室可被加热至适宜的反应温度，以便使样本获得最佳的处理效果。
在混合室中样本经相应的试剂处理后制备得到待测样本。待测样本通过管路组件输送到检测单元。在该实施方式中，待测样本被输送到光学检测单元。
在该实施方式中，光源、光学流动室、检测孔以及光学检测器均属于光学检测单元的部件。
光源用于将光束对准与所述混合室相连通的所述光学流动室的检测孔。光源通常为激光器，其波长可以是蓝光范围也可以是红光范围。根据本公开的方法，可选择适用于不同波长光源的荧光标记的抗体中的荧光标记物，配合所用血细胞分析仪配置的试剂(如含有第一荧光染料的第三试剂和/或含有第二荧光染料的第五试剂)进行上述样本分析。
光学检测器用于检测逐一通过检测孔的待测试样中粒子的光散射信号和荧光信号。其中，光散射信号包括由设置在光轴上的前向散射光信号收集装置收集的前向光散射强度信号，和由设置在光轴侧边的侧向散射光信号收集装置收集的侧向光散射强度信号。此外，光学检测器还包括设置在光轴侧边的荧光信号收集装置收集荧光强度信号。根据样本的不同处理，荧光信号收集装置可以收集来自已经结合到细胞膜表面的荧光标记的抗体发出的抗体荧光强度信号，和/或收集来自进入到细胞内部与核酸或蛋白质结合的荧光染料(例如第一或第二荧光染料)发出的荧光强度信号。
光学检测器检测到粒子的光散射信号和荧光信号被传输到处理器，进行分析。
所述处理器与所述光学检测器可操作地连接，当所述非暂时性计算机可读存储介质中的计算机应用程序被所述处理器执行时，控制试剂与子样本在混合室中的混合时间，且所述处理器根据接收到的光散射信号和体荧光信号获得相应的粒子信息，并针对各检测方法(例如上述实施方式中任意一种原始细胞检测方法和/或常规检测方法)执行获取粒子信息的各步骤。获取粒子信息的各步骤与前述方法中相同，在此不再赘述。
根据本公开的另一种实施方式，所述细胞分析仪还可包括血红蛋白检测单元和阻抗计数检测单元。
其中血红蛋白检测单元可用于血红蛋白检测。阻抗计数检测单元可用于红细胞和/或血小板的检测。
以下参考图10～12，说明本公开具体实施方式提供的一种血细胞分析仪。本领域技术人员应知，用于本公开的血细胞分析仪不限于以下详述的实施方式。
该实施方式中，血细胞分析仪包括吸样装置100、光学通道混合室200、光学检测单元300、血红蛋白检测单元400、阻抗计数检测单元500、第一试剂供应装置600、第二试剂供应装置700、输送管路组件800、流体动力装置900、废液池1000、稀释液提供装置1100、以及处理器和编程有计算机应用程序的非暂时性计算机可读存储介质(未示出)，吸样装置100、光学通道混合室200、光学检测单元300、血红蛋白检测单元400、阻抗计 数检测单元500、第一试剂供应装置600、第二试剂供应装置700、废液池1000、稀释液提供装置1100和流体动力装置900通过输送管路组件800连接形成一个液路系统。
本实施方式中，流体动力装置900包括第一注射器910和第二注射器920，第一注射器900的量程小于第二注射器920的量程。其中，第一注射器910复用于为采样、分血(分样)、推样(包括光学通道推样和鞘流阻抗推样)提供动力。第二注射器920复用于为样本稀释、样本流经部件清洗、样本准备、阻抗计数检测单元500充灌、光学检测推鞘液提供动力。本实施方式中，流体动力装置900只有两支注射器高度复用，能够达到降低仪器的成本及体积的目的。
优选地，第一注射器910和第二注射器920分别采用独立的动力机构驱动。具体地，流体动力装置900还包括用于驱动第一注射器910动作的第一电机(图未示)和用于驱动第二注射器920动作的第二电机(图未示)，第一电机与第二电机为两个相互独立设置的电机。第二电机的步长优选大于第一电机的步长。
光学检测单元300包括流动室310和光学检测元件320，流动室310具有检测区、稀释液入口d、样本入口e和第一出口f，稀释液入口d、样本入口e和第一出口f分别与检测区连通。检测区用于供光学检测样本在稀释液裹挟下通过。光学检测元件320可包括光源、设置在光轴上的前向散射光信号收集装置、设置在光轴侧边的侧向散射光信号收集装置和荧光信号收集装置。
光学通道混合室200用于为待测样本和试剂提供反应场所，以制备得到光学待测试样。
本实施方式中，光学通道混合室200包括第一混合室210和第二混合室220，第一混合室210和第二混合室220为两个相互独立的混合室，即光学通道混合室200的数量为两个。其中，第一混合室210用于为例如第一子样本和检测试剂(例如第一和第二试剂，或第一、第二和第三试剂)提供反应场所，以制备得到第一待测试样。第二混合室220用于为第二子样本和检测试剂(如第三和第四试剂)提供反应场所，以制备得到第二待测试样。本实施方式中，以两个子样本在两个相互独立的混合室内进行制备为例进行说明。当然，根据前述方法，本公开一些实施方式的血细胞分析仪还可包括第三混合室，用于第三子样本的处理。具有多个混合室的分析仪，一方面可以避免共用混合室时因试剂残留产生交叉污染的问题产生；另一方面可以使得多个子样本能够同时制备，利于提高检测效率。
输送管路组件800包括光学样本准备管路810、采样输送管820、第一连接管路830、第二连接管路840、第三连接管路850、第四连接管路860、第五连接管路870、第一接头880、第二接头890、第三接头8100、第四接头8110、第一稀释液输送管路8120、第三稀释液输送管路8140、第四稀释液输送管路8150、第一输送管路8160、阻抗计数样本准备管路8170、第二输送管路8180和第七接头8190、第二稀释液输送管路8200、第三输送管路8210、第一排液管路8220、第二排液管路8230、第三排液管路8240、第四排液管路8250和第五排液管路8260。其中，光学样本准备管路810分别与样本入口e、光学通道混合室200、第一注射器910、第二注射器920连接，第一稀释液输送管路8120分别与稀释液入口d、稀释液提供装置1100连接，第二稀释液输送管路8200分别与稀释液提供装置1100、血红蛋白检测单元400、第二注射器920连接，第三稀释液输送管路8140分别与稀释液提供装置1100、阻抗计数检测单元500、第二注射器920连接。
稀释液提供装置1100用于提供稀释液。第二注射器920通过第一稀释液输送管路8120分别连接稀释液提供装置1100和稀释液入口d，以用于驱动稀释液从稀释液提供装置1100输送至流动室310；第二注射器920与光学样本准备管路810连接，以用于驱动光学检测样本从光学通道混合室200内输送至光学样本准备管路810内；第二注射 器920还通过第二稀释液输送管路8200分别连接稀释液提供装置1100和血红蛋白检测单元400，以用于驱动稀释液从稀释液提供装置1100输送至血红蛋白检测单元400；第二注射器920还通过第三稀释液输送管路8140分别连接稀释液提供装置1100和阻抗计数检测检测单元500，以用于驱动稀释液从稀释液提供装置1100输送至阻抗计数检测检测单元500。
吸样装置100用于采集待测样本。本实施方式中，待测样本为血液样本，当然了，作为替代的实施方案，待测样本也可为体液样本。采样输送管820连接于第一注射器910与吸样装置100之间。第一注射器910与吸样装置100连接以用于驱动吸样装置100进行吸取待测样本(即采样)和驱动吸样装置100内的待测样本分别输送至血红蛋白检测单元400和光学通道混合室200内(即分血)进行反应。吸样装置100可为采样针或者采样吸管等。血细胞分析仪还包括用于驱动吸样装置100运动的动力元件(图未示)，吸样装置100可在动力元件的驱动下移动至样本存放容器(例如试管等)进行采样，然后在动力元件的驱动下分别移动至光学通道混合室200和血红蛋白检测单元400，并在第一注射器910的驱动下进行分血。
第一注射器910通过光学样本准备管路810连接样本入口e，以用于驱动光学检测样本从光学样本准备管路810内输送至流动室310内。
在原始细胞检测时，第二注射器920用于驱动第一待测试样从第一混合室210内输送至光学样本准备管路810和驱动稀释液输送至流动室310内；第一注射器910用于驱动第一待测试样从光学样本准备管路810内输送至流动室310内；光学检测元件320用于对由稀释液裹挟通过检测区的第一待测试样进行原始细胞检测，并根据散射光信号和抗体荧光信号获得原始细胞检测结果。
在白细胞分类检测时，第一注射器910用于驱动第二待测试样从光学样本准备管路810内输送至流动室310内和驱动稀释液输送至流动室310内，第二注射器920用于驱动第二待测试样从第二混合室200内输送至光学样本准备管路810内；光学检测元件320用于对由鞘液裹挟通过检测区的第二待测试样进行白细胞分类检测。
第一试剂供应装置600用于为光学通道混合室200提供试剂。本实施方式中，第一试剂供应装置600包括第一定量泵610、第二定量泵620、第三定量泵630和第四定量泵640。
第一定量泵610与第一混合室210连接以用于为第一混合室210提供第一试剂；第二定量泵620与第一混合室210连接以用于为第一混合室210提供第二试剂。作为本实施方式的一较佳实施方案，第一试剂和第二试剂中分别含有溶血剂和荧光标记的抗体，荧光标记的抗体试剂用于在原始细胞检测时使得光学检测单元300能够获取抗体荧光信号，溶血剂主要用于使红细胞破碎。当然了，具体应用中，作为替代的实施方案，第一试剂和第二试剂也可以由一种同时具有溶血剂和荧光标记的抗体的试剂代替，该替代的实施方案中，与第一混合室210连接的定量泵只有一个，例如第一定量泵610。
第三定量泵630与第二混合室220连接以用于为第二混合室220提供第三试剂；第四定量泵640与第二混合室220连接以用于为第二混合室220提供第四试剂。第三试剂和第四试剂分别含有第二溶血剂和第一荧光染料。当然了，具体应用中，作为替代的实施方案，溶血剂和第一荧光染料也可以包含在单一一种试剂中作为代替，该替代的实施方案中，与第二混合室220连接的定量泵只有一个，例如第三定量泵630。
血红蛋白检测单元400用于为待测样本和例如第七试剂提供反应场所以制备得到血红蛋白检测样本、为待测样本和稀释液提供反应场所以制备得到阻抗计数检测样本以及用于对血红蛋白检测样本进行血红蛋白浓度检测。本实施方式中，血红蛋白检测单元400除了作为血红蛋白检测样本的制备单元和检测单元使用外，还作为阻抗计数检测样本制备单元使用，即阻抗计数检测样本制备单元与血红蛋白检测单元400一体设置。具 体操作时，可先在血红蛋白检测单元400中加入待测样本和稀释液反应，制得阻抗计数检测样本，然后将部分阻抗计数检测样本抽走；然后再向血红蛋白检测单元400中加入第七试剂制备血红蛋白检测样本。
第二试剂供应装置700用于为血红蛋白检测单元400提供第七试剂，其包括与血红蛋白检测单元400连接的第五定量泵710。第五试剂例如为能够溶解血液样本中的红细胞、释放红细胞中的血红蛋白并将血红蛋白转化为高铁血红蛋白的溶血剂。在替代的实施方案中，白细胞分类检测中用的试剂与血红蛋白检测中用试剂也可为相同的溶血剂，即用于给第二混合室220添加试剂的定量泵和用于为血红蛋白检测单元400添加试剂的定量泵为同一部件。
光学样本准备管路810设置于流动室310与光学通道混合室200之间。第二注射器920与光学样本准备管路810连接以用于驱动光学检测样本从光学通道混合室200内输送至光学样本准备管路810内，第一注射器910通过光学样本准备管路810连接样本入口e以用于驱动光学检测样本从光学样本准备管路810内输送至流动室310内。
本实施方式中，光学样本准备管路810包括第一样本准备管路811和第二样本准备管路812，即光学样本准备管路810的数量为两个，第一样本准备管路811和第二样本准备管路812为两条相互独立的管路。第一样本准备管路811分别与第一混合室210、流动室310、第一注射器910和第二注射器920连接，第二样本准备管路812分别与第二混合室220、流动室310、第二混合室220、第一注射器910和第二注射器920连接。在原始细胞检测时，第二注射器920用于驱动第一待测试样从第一混合室210内驱动输送至第一样本准备管路811内和驱动鞘液输送至流动室310内，第一注射器910用于驱动第一待测试样从第一样本准备管路811输送至流动室310内。在进行白细胞分类检测时，第二注射器920驱动第二待测试样从第二混合室220内驱动输送至第二样本准备管路812内和驱动鞘液输送至流动室310内，第一注射器910用于驱动第二待测试样从第二样本准备管路812输送至流动室310内。本实施方式中，原始细胞检测和白细胞分类检测分别采用独立的混合室和独立的样本准备管路，利于减少交叉污染。当然了，具体应用中，光学通道混合室200包括的混合室数量不限于两个。
第一样本准备管路811的一端与流动室310连接、另一端通过第一连接管路830连接第一混合室210，第二样本准备管路812的一端与流动室310连接、另一端通过第三连接管路850连接第二混合室220，第二注射器920通过第二连接管路840分别连接第一样本准备管路811和第二样本准备管路812,第一注射器910通过第四连接管路860连接第一样本准备管路811、并通过第五连接管路870连接第二样本准备管路812连接。
第一连接管路830上设有第一可控阀831，第二连接管路840上设有第二可控阀841，第三连接管路850上设有第三可控阀851。第一控制阀8121、第二可控阀841和第一可控阀831打开时，第二注射器920可以向第一混合室210驱动输送稀释液；第一控制阀8121关闭，第二可控阀841和第一可控阀831打开时，第二注射器920可以驱动原始细胞检测样本从第一混合室210输送至第一样本准备管路811；第一控制阀8121、第二可控阀841和第三可控阀851打开时，第二注射器920可以向第二混合室220驱动输送稀释液；第一控制阀8121关闭，第二可控阀841和第三可控阀851打开时，第二注射器920可以驱动白细胞分类检测样本从第二混合室220输送至第二样本准备管路812。
第四连接管路860的一端连接第一注射器910、另一端连接第一样本准备管路811，第四连接管路860上设有第六控制阀861，第五连接管路870的一端连接第六控制阀861、另一端连接第二样本准备管路812。第六控制阀861可以控制第一注射器910分别切换连接第一样本准备管路811和第二样本准备管路812。当然了，作为替代的实施方案，第四连接管路860和第五连接管路870也可通过两个相互独立的控制阀分别连接第一注射器910。
优选地，本实施方式中，第四连接管路860上还设有第五可控阀862，第五可控阀862位于第一注射器910与第四可控阀861之间，吸样装置100通过第五可控阀862连接第一注射器910。采样输送管820的一端连接吸样装置100、另一端连接第五可控阀862。第五可控阀862可以控制第一注射器910分别切换连接吸样装置100和样本准备管路。当然了，作为替代的实施方案，样本准备管路和吸样装置100也可通过两个相互独立的可控阀分别连接第一注射器910。
第一接头880具有相互连通的第一接口a、第二接口b和第三接口c，第一接口a与流动室310的样本入口e连接，第三接口c通过第二连接管路840与第二注射器920连接；第一样本准备管路811和第二样本准备管路812中的一者连接于第二接口b与光学通道混合室200之间，第一样本准备管路811和第二样本准备管路812中的另一者一端与光学通道混合室200连接、另一端分别连接第二注射器920和第三接口c，第四连接管路860通过第三接头8100连接第一样本准备管路811和第一连接管路830，第五连接管路870通过第四接头8110连接第二样本准备管路812和第三连接管路850。第三接头8100和第四接头8110都优选为三通接头，即第一注射器910通过两个三通接头分别与两条光学样本准备管路810连接。
本实施方式中，第二连接管路840还通过第一稀释液输送管路8120分别连接第二注射器920和稀释液提供装置1100，即：第二连接管路840的一端分别与第一样本准备管路811、第二样本准备管路812连接，第二连接管路840的另一端与第一稀释液输送管路8120连接。第二注射器920还用于驱动稀释液从稀释液提供装置1100输送至光学样本准备管路810和光学通道混合室200，这样，第二注射器920可为稀释液清洗第一样本准备管路811、第二样本准备管路812、第一混合室210和第二混合室220提供动力。
第一稀释液输送管路8120的一端连接流动室310的稀释液入口d、另一端连接第二注射器920，稀释液提供装置1100和第二连接管路840分别连接于第一稀释液输送管路8120。第一稀释液输送管路8120上设有第一控制阀8121和第二控制阀8122，第一控制阀8121靠近第二注射器920设置，第二控制阀8122靠近稀释液入口d设置，稀释液提供装置1100连接于第一控制阀8121。第一控制阀8121和第二控制阀8122可以控制第一稀释液输送管路8120的开启和关闭。具体应用中，当打开第一控制阀8121和第二控制阀8122时，第二注射器920可以向流动室310内推送鞘液；当关闭第一控制阀8121和第二控制阀8122时，第二注射器920不能向流动室310内推送鞘液。
第二注射器920通过第四稀释液输送管路8150分别连接稀释液提供装置1100和第一注射器910，以用于驱动稀释液从稀释液提供装置1100输送至第一注射器910。具体地，第四稀释液输送管路8150连接于第一稀释液输送管路8120与第一注射器910之间。第四稀释液输送管路8150上设有第五控制阀8151，打开第一控制阀8121和第五控制阀8151时，第二注射器920可以驱动稀释液输送至第一注射器910，以便于清洗第一注射器910、吸样装置100和采样输送管820。
阻抗计数检测单元500用于对阻抗计数检测样本进行阻抗计数检测，阻抗计数检测可包括红细胞数目检测和/或血小板计数检测。阻抗计数检测检测样本可由待测样本和稀释液反应制得。
阻抗计数检测单元500包括阻抗计数检测装置510、稀释液储液池520、正压动力源530、压力传感器540和第五稀释液输送管路550，第五稀释液输送管路550上设有第六控制阀551，正压动力源530、压力传感器540分别与稀释液储液池520，其中，正压动力源530通过气阀570连接稀释液储液池520。稀释液储液池520通过第五稀释液输送管路550连接阻抗计数检测装置510、并通过第三稀释液输送管路8140连接第二注射器920和稀释液提供装置1100；阻抗计数样本准备管路8170的一端连接阻抗计 数检测装置510、并通过第一输送管路8160连接第二注射器920，阻抗计数样本准备管路8170的另一端通过第二输送管路8180连接血红蛋白检测单元400、并通过第三输送管路8210连接第一注射器910；第二注射器920还用于驱动稀释液从稀释液提供装置1100输送至稀释液储液池520和驱动阻抗计数检测样本从血红蛋白检测单元400输送至阻抗计数样本准备管路8170，正压动力源530用于驱动稀释液从稀释液储液池520输送至阻抗计数检测装置520、以使阻抗计数检测样本在稀释液的裹挟下在阻抗计数检测装置520内流动；第一注射器910还用于驱动阻抗计数检测样本从阻抗计数样本准备管路8170输送至阻抗计数检测装置510。
具体地，第三稀释液输送管路8140的一端与第一稀释液输送管路8120连接、另一端与稀释液储液池520连接；第一输送管路8160的一端与第三稀释液输送管路8140连接、另一端分别与阻抗计数样本准备管路8170和阻抗计数检测装置510连接。第一输送管路8160上设有第七控制阀8161，第三稀释液输送管路8140上设有第四控制阀8141，第五稀释液输送管路550上设有第六控制阀551。当打开第一控制阀8121和第四控制阀8141时，第二注射器920可以驱动稀释液向稀释液储液池520输送以实现充灌；当打开气阀570和第六控制阀551时，正压动力源530可以驱动鞘液从稀释液储液池520内输送至阻抗计数检测装置510内。本实施方式，第三稀释液输送管路8140是通过第一稀释液输送管路8120间接连接稀释液提供装置1100和第二注射器920的；当然了，具体应用中，作为替代的实施方案，也可以改为第三稀释液输送管路8140直接连接稀释液提供装置1100和第二注射器920的方案。
阻抗计数样本准备管路8170的一端通过第七接头8190分别连接阻抗计数检测装置510和第一输送管路8160，阻抗计数样本准备管路8170的另一端分别连接第二输送管路8180和第三输送管路8210。第二输送管路8180连接于阻抗计数样本准备管路8170与血红蛋白检测单元400之间。第二输送管路8180上设有第八控制阀8181。当第七控制阀8161和第八控制阀8181打开时，第二注射器920可以驱动阻抗计数检测样本从血红蛋白检测单元400输送至阻抗计数样本准备管路8170内。
阻抗计数检测装置510具有前池(图未标示)、后池(图未标示)、宝石孔(图未标示)、第二入口h、第一入口i和第二出口g，第二入口h、第一入口i分别与前池连通，宝石孔用于连通前池与后池，第二出口g与后池连通。第二入口h与第五稀释液输送管路550连接，第一入口i通过第七接头8190分别连接第一输送管路8160和阻抗计数样本准备管路8170，第二出口g与废液池1000连接。
第二稀释液输送管路8200连接于第一稀释液输送管路8120与血红蛋白检测单元400之间，阻抗计数样本准备管路8170分别与第三输送管路8210的一端和第二输送管路8180的一端连接，第三输送管路8210的另一端连接第一注射器910，第二输送管路8180的另一端连接第二稀释液输送管路8200。第二稀释液输送管路8200上设有第三控制阀8201，第五定量泵710与第二稀释液输送管路8200连接，且第五定量泵710、第二输送管路8180在第二稀释液输送管路8200上的连接点都位于第三控制阀8201与血红蛋白检测单元400之间。打开第一控制阀8121和第三控制阀8201时，第二注射器920可以向血红蛋白检测单元400输送稀释液。本实施方式，第二稀释液输送管路8200是通过第一稀释液输送管路8120间接连接稀释液提供装置1100和第二注射器920的；当然了，具体应用中，作为替代的实施方案，也可以改为第二稀释液输送管路8200直接连接稀释液提供装置1100和第二注射器920的方案。
第三输送管路8210上设有第九控制阀8211，第五可控阀862位于第九控制阀8211与第一注射器910之间。第四连接管路860连接第九控制阀8211。第五可控阀862可以控制第一注射器910与采样输送管820之间通道的通断以及控制第一注射器910与第九控制阀8211之间通道的通断，即通过调控第五可控阀862可以切换至第一注射器910 与采样输送管820连通的状态和切换第一注射器910与样本准备管路(包括光学样本准备管路810和阻抗计数样本准备管路8170)连通的状态。通过第九控制阀8211和第五可控阀862的配合，可以控制第一注射器910与阻抗计数样本准备管路8170之间通道的通断；通过第六控制阀861、第九控制阀8211和第五可控阀862的配合，可以控制第一注射器910与第一样本准备管路811、第二样本准备管路812之间通道的通断。
本实施方式中，第一注射器910通过第九控制阀8211分别连接阻抗计数样本准备管路8170和光学样本准备管路810。当然了，具体应用中，作为替代的实施方案，光学样本准备管路810和阻抗计数样本准备管路8170也可分别通过相互独立的控制阀连接第一注射器910。
第一排液管路8220连接于流动室310的第一出口f与废液池1000之间，第一排液管路8220上设有第一排液控制阀8221。在原始细胞检测和白细胞分类检测时，打开第一排液控制阀8221，可将通过检测区从流动室310内流出的网织红细胞检测样本和稀释液或者白细胞分类检测样本和稀释液输送到废液池1000。
第二排液管路8230连接于第一混合室210与废液池1000之间，第二排液管路8230上设有第二排液控制阀8231。打开第二排液控制阀8231，可以排空第一混合室210，以便于在清洗第一混合室210时，清洗废液可以排放到废液池1000。
第三排液管路8240连接于第二混合室220与废液池1000之间，第三排液管路8240上设有第三排液控制阀8241。打开第三排液控制阀8241，可以排空第二混合室220，以便于在清洗第二混合室220时，清洗废液可以排放到废液池1000。
第四排液管路8250连接于血红蛋白检测单元400与废液池1000之间，第四排液管路8250上设有第四排液控制阀8251。打开第四排液控制阀8251，可以排空血红蛋白检测单元400，以便于在清洗血红蛋白检测单元400时，清洗废液可以排放到废液池1000。
第五排液管路8260连接于阻抗计数检测装置510的第二出口g与废液池1000之间。在阻抗计数检测时，从阻抗计数检测装置510的第二出口g流出的阻抗计数检测样本和鞘液可以排放到废液池1000。
需要说明的是，废液池1000可以只有一个，即第一排液管路8220、第二排液管路8230、第三排液管路8240、第四排液管路8250和第五排液管路8260与同一个废液池1000连接；当然了，作为替代的实施方案，废液池1000也可以设有两个以上，第一排液管路8220、第二排液管路8230、第三排液管路8240、第四排液管路8250和第五排液管路8260中的至少两者分别连接至不同的废液池1000。
以下参考图17～18，说明本公开另一具体实施方式的样本分析仪2100。
本实施方式提供的样本分析仪2100，保护侧重点在于原始细胞检测与血常规检测一体机的实现方案。
具体地，本实施方式提供的样本分析仪2100，包括样本分配装置2111、光学检测装置2112、第一反应装置2113、第二反应装置2116、第一试样输送组件2114和控制器2120；样本分配装置2111用于从样本容器2200中吸取样本，并将吸取的至少部分样本分配至第一反应装置2113和/或第二反应装置2116；第一反应装置2113至少用于为样本与抗体试剂、第一溶血试剂提供反应场所以制备形成第一待测试样，其中，抗体试剂中的抗体能够与样本中原始细胞的表面抗原相结合；第二反应装置2116至少用于为样本与第一类试剂提供反应场所以制备形成第二待测试样；第一试样输送组件2114用于将第一待测试样从第一反应装置2113输送至光学检测组件，且用于将第二待测试样从第二反应装置2116输送至光学检测组件；光学检测组件包括流动室、光发射部件和光接收部件，流动室用于供第一待测试样或第二待测试样在鞘液的裹挟下通过，光发射部件用于朝向流动室中的第一待测试样或第二待测试样发射光线，光接收部件用于接收光发射部件发射的光线经第一待测试样产生的第一光学信号以及用于接收光发射部 件发射的光线经第而待测试样产生的第二光学信号；控制器2120被配置为：根据光接收部件反馈的第一光学信号，解析得到第一待测试样中的第一粒子信息，第一粒子信息包括原始细胞信息；根据光接收部件反馈的第二光学信号，解析得到第二待测试样中的第二粒子信息，第二粒子信息包括中性粒细胞信息、嗜酸性粒细胞信息、单核细胞信息和淋巴细胞信息。本实施方式中，为原始细胞和白细胞分类分别设置独立的场所，利于使得白细胞检测中的反应，与原始细胞检测中的反应可以同时进行，从而利于提高检测通量。白细胞分类计数与原始细胞计数检测共用一个光学检测装置2112，成本低。本实施方式的原始细胞检测中，由于不需要人工手动分配样本，故，故，具有自动化程度高、操作简单、省时省力、效率高的优势，且消除了人为定量误差的隐患。此外，本实施方式通过样本分析仪2100设置固定的第一反应装置2113进行原始细胞检测中样本与抗体试剂的反应，不需要在样本分析仪2100外设置空白的样本容器2200进行样本与抗体试剂反应，减小了样本容器2200不必要的消耗。
作为一种实施方式，样本分析仪2100还包括外壳2130，光学检测装置2112、第一反应装置2113和第二反应装置2116都位于外壳2130内。本实施方案中，原始细胞检测的抗体反应在样本分析仪2100内部进行，不需要在样本分析仪2100外设置空白的样本容器2200进行样本与抗体试剂反应，减小了样本容器2200不必要的消耗。
作为一种实施方式，样本分析仪2100还包括试剂存储装置2115，试剂存储装置2115至少用于存放第一试剂容器，第一试剂容器用于装载抗体试剂，试剂存储装置2115设于外壳2130内。本实施方案中，抗体试剂也存放于样本分析仪2100内。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的样本分配过程中，控制样本分配装置2111分配至第一反应装置2113的样本量小于或等于60μL。通过对原始细胞检测的待测试样制备方式进行优化设计，并对原始细胞检测结果的解析方式进行优化设计，从而可以在原始细胞检测结果准确性的前提下，使得单个原始细胞检测项目所用的样本量可以减小至小于或等于60μL，相对于传统流式细胞分析仪中需要至少100μL的样本而言，大幅度降低了原始细胞检测中样本的用量。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的样本分配过程中，控制样本分配装置2111分配至第一反应装置2113的样本量在50μL±10μL之间。可以理解的，如果检测所用的样本量过大，则需要从患者身上采集的样本量相对也比较大；而如果检测所用的样本量过小，则又不利于保证检测结果的准确性。本实施方案中，将单个原始细胞检测项目所用的样本量控制在50μL左右，既利于减小样本量，又利于保证原始细胞检测结果的准确性。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的样本分配过程中，控制样本分配装置2111分配至第一反应装置2113的样本量为50uL。本实施方案中，将单个原始细胞检测项目所用的样本量控制在50μL。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的抗体试剂分配过程中，控制分配至第一反应装置2113的抗体试剂的量在5μL±2μL之间。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的抗体试剂分配过程中，控制分配至第一反应装置2113的抗体试剂的量小于或等于5μL。本实施方案，通过对原始细胞检测的待测试样制备方式进行优化设计，并对原始细胞检测结果的解析方式进行优化设计，从而可以采用极少的抗体试剂量，即可进行原始细胞的准确检测，利于减少试剂成本和反应时间。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的抗体试剂分配过程中，控制分配至第一反应装置2113的抗体试剂的量小于5μL。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的检测过程中，控制样本与抗体试剂在第一反应装置2113的反应时间小于或等于90s。本实施 方案，通过对原始细胞检测的待测试样制备方式进行优化设计，并对原始细胞检测结果的解析方式进行优化设计，从而可以在保证检测结果准确性的前提下，减小样本与抗体试剂的反应时间，进而利于提高原始细胞的检测效率。
作为一种实施方式，控制器2120还被配置为：在单个原始细胞检测项目的检测过程中，控制样本与抗体试剂在第一反应装置2113的反应时间小于或等于60s。
作为一种实施方式，抗体试剂中包含经荧光标记、且能够与原始细胞表面抗原相结合的抗体，抗体被荧光染料标记。第一光学信号包括由光发射部件发射的光线照射至第一待测试样产生的第一侧向散射光信号和由光发射部件发射的光线照射至第一待测试样经荧光标记产生的第一荧光信号。第一侧向散射光信号和第一荧光信号可以将原始细胞与成熟的白细胞区分开。具体地，本实施方案，利用经荧光标记的抗体与不同细胞表面抗原结合程度的不同，得到不同的荧光信号，同时结合反映细胞内部结构复杂度的侧向散射光，能够直接使原始细胞区分于其它白细胞，从而利于实现对原始细胞的准确计算。
作为一种实施方式，抗体试剂为CD45试剂，CD45试剂包含经荧光标记、且能够与原始细胞表面抗原相结合的抗体，且该抗体可以与原始细胞的表面抗原进行特异性结合，以利于通过荧光信号区分原始细胞与白细胞。在本申请之前，没有发现采用CD45与样本进行抗体抗原反应进行原始细胞检测的方案；而本申请的发明人，发现采用CD45与样本进行抗体抗原反应，再进行溶血反应，可以极大程度地减小原始细胞检测所用的样本量和试剂量，且可以缩短样本的反应时间。
上述方案中，荧光染料与抗体为一体设置，即荧光试剂与抗体合为一种试剂，第一反应液由样本与单一的一种试剂反应制成。当然，具体应用中，作为替代的实施方案，荧光染料与抗体也可以分开设于两种不同的试剂中，即用于与样本反应制成第一反应液的试剂也可以是两种试剂，一种是抗体试剂，另一种是荧光试剂。该替代的实施方案中，抗体试剂中包含未经荧光标记、但能够与原始细胞表面抗原相结合的抗体，第一反应装置2113用于为样本与抗体试剂、第一荧光试剂提供反应场所以制备形成第一反应液，第一光学信号包括由光发射部件发射的光线照射至第一待测试样产生的第一侧向散射光信号和由光发射部件发射的光线照射至第一待测试样经第一荧光试剂产生的第一荧光信号。
作为一种实施方式，第一反应装置2113用于为第一反应液与第一溶血试剂、第二荧光试剂提供反应场所以制备形成第一待测试样；第一光学信号还包括由光发射部件发射的光线照射至第一待测试样经第二荧光试剂产生的第二荧光信号。控制器2120根据光接收部件反馈的第一光学信号，解析得到第一待测试样中的第一粒子信息：根据第一侧向散射光信号和第一荧光信号解析得到第一原始细胞信息和第一血影区域信息，根据第二荧光信号解析得到第二血影区域信息；将第一原始细胞信息和第一血影区域信息，扣减第二血影区域信息，得到第一待测试样的原始细胞信息。本实施方案中，通过在溶血反应中，进一步增加第二荧光试剂，从而利于更好地标记血影区域；通过扣减血影区域对原始细胞检测的影响，可以得到更加准确的原始细胞信息。需要说明的是，控制器2120根据第一侧向散射光信号和第一荧光信号，除了可以解析得到第一原始细胞信息和第一血影区域信息外，还可以得到区分于原始细胞的成熟白细胞信息。
作为一种实施方式，样本分配装置2111还用于从第一试剂容器中吸取抗体试剂，并将吸取的抗体试剂分配至第一反应装置2113。本实施方案中，样本分配装置2111分时复用于样本的分配和抗体试剂的分配，可以在不额外增加分配装置的前提下，实现不需要人工手动分配抗体试剂的效果，利于进一步提高样本分析仪2100的自动化程度，且消除了人为定量抗体试剂误差的隐患。当然，具体应用中，抗体试剂不限于采用样本分配装置2111进行分配，例如，作为一种替代的实施方案，样本分析仪2100还包括第 一试剂分配装置，第一试剂分配装置用于从第一试剂容器中吸取抗体试剂，并将吸取的抗体试剂分配至第一反应装置2113，该替代方案中，采用不同于样本分配装置2111的第一试剂分配装置进行分配抗体试剂。
作为一种实施方式，第二反应装置2116包括第三反应池，且包括第四反应池和第五反应池中的至少一者，第三反应池用于为样本与第一类试剂提供反应场所以制备形成第二待测试样，第四反应池用于为样本与第二类试剂提供反应场所以制备形成第三待测试样，第五反应池用于为样本与第三类试剂提供反应场所以制备形成第四待测试样。第二待测试样用于白细胞四分类计数，即：第三反应池为DIFF反应通道，第二待测试样用于检测样本中的中性粒细胞信息、嗜酸性粒细胞信息、单核细胞信息和淋巴细胞信息。第三待测试样用于白细胞分类中的嗜碱性粒细胞计数，即：第四反应池为WNB反应通道，第三待测试样用于检测样本中的嗜碱性粒细胞信息。结合第二待测试样和第三待测试样的检测结果，即实现了白细胞的五分类计数。第四待测试样用于网织红细胞计数，即：第五反应池为RET反应通道，第四待测试样用于检测样本中的网织红细胞信息。
作为一种实施方式，第二反应装置2116包括第三反应池、第四反应池和第五反应池，样本分析仪2100除了可以进行原始细胞计数外，还可以实现白细胞分类计数和网织红细胞计数。当然，具体应用中，第二反应装置2116的设置方式不限于此，例如，作为一种替代的实施方案，第二反应装置2116也可以只用于进行白细胞分类计数，且白细胞分类计数既可以是五分类计数，也可以是四分类计数，或者三分类计数；或者，作为另一种替代的实施方案，第二反应装置116也可以只包括一个反应池，该反应池既可以用于白细胞分类计数的待测试样制备，也可以用于网织红细胞计数的待测试样制备。
作为一种实施方式，第一试样输送组件2114还用于将第二待测试样或第三待测试样或第四待测试样从第二反应装置2116输送至流动室。当第二反应装置2116包括第三反应池、第四反应池和第五反应池时，第一试样输送组件2114分时复用于：将第一待测试样从第一反应池21131输送至流动室，将第二待测试样从第三反应池输送至流动室，将第三待测试样从第四反应池输送至流动室，将第四待测试样从第五反应池输送至流动室。本实施方案中，原始细胞通道与血常规光学检测通道共用一个试样输送组件将待测试样输送至流动室，具有结构简单、成本低的优点；当然，具体应用中，作为替代的实施方案，原始细胞通道与血常规光学检测通道也可以分别采用不同试样输送组件将待测试样输送至流动室。
作为一种实施方式，流动室还用于供第二待测试样或第三待测试样或第四待测试样在鞘液的裹挟下通过。当第二反应装置2116包括第三反应池、第四反应池和第五反应池时，流动室分时复用于：供第一待测试样在鞘液的裹挟下通过，供第二待测试样在鞘液的裹挟下通过，供第三待测试样在鞘液的裹挟下通过，供第四待测试样在鞘液的裹挟下通过。
作为一种实施方式，光发射部件还用于朝向流动室中的第二待测试样或第三待测试样或第四待测试样发射光线。当第二反应装置2116包括第三反应池、第四反应池和第五反应池时，光发射部件分时复用于：朝向流动室中的第一待测试样发射光线，朝向流动室中的第二待测试样发射光线，朝向流动室中的第三待测试样发射光线，朝向流动室中的第四待测试样发射光线。
作为一种实施方式，光接收部件还用于接收光发射部件发射的光线经第二待测试样产生的第二光学信号、或经第三待测试样产生的第三光学信号、或经第四待测试样产生的第四光学信号。当第二反应装置2116包括第三反应池、第四反应池和第五反应池时，光接收部件分时复用于：接收光发射部件发射的光线经第一待测试样产生的第一光学信号，接收光发射部件发射的光线经第二待测试样产生的第二光学信号，或经第三待测试样产生的第三光学信号，或经第四待测试样产生的第四光学信号。
作为一种实施方式，第一光学信号包括由光发射部件发射的光线照射至第一待测试样产生的第一侧向散射光信号、由光发射部件发射的光线照射至第一待测试样经荧光标记或第一荧光试剂产生的第一荧光信号以及由光发射部件发射的光线照射至第一待测试样经第二荧光试剂产生的第二荧光信号。
作为一种实施方式，第一类试剂包括第三荧光试剂和第二溶血试剂，第二光学信号包括由光发射部件发射的光线照射至第二待测试样产生的第二侧向散射光信号和第三荧光信号。
作为一种实施方式，第二类试剂包括第四荧光试剂和第三溶血试剂，第三光学信号包括由光发射部件发射的光线照射至第三待测试样产生的第三侧向散射光信号和第四荧光信号。
作为一种实施方式，第三类试剂包括第五荧光试剂，第四光学信号包括由光发射部件发射的光线照射至第四待测试样产生的第四侧向散射光信号和第五荧光信号。
作为一种实施方式，光接收部件包括侧向散射光探测器、前向散射光探测器、第一荧光探测器和第二荧光探测器，侧向散射光探测器用于接收第一侧向散射光信号、第二侧向散射光信号、第三侧向散射光信号和第四侧向散射光信号，前向散射光探测器用于接收由光发射部件发射的光线照射至待测试样产生的前向散射光信号，第一荧光探测器用于接收第一荧光信号，第二荧光探测器用于接收第二荧光信号、第三荧光信号、第四荧光信号和第五荧光信号。
作为一种实施方式，控制器2120还被配置为根据光接收部件的反馈信息执行如下的至少一个解析动作：根据光接收部件反馈的第二光学信号，解析得到第二待测试样中的第二粒子信息，第二粒子信息包括中性粒细胞信息、嗜酸性粒细胞信息、单核细胞信息和淋巴细胞信息；根据光接收部件反馈的第三光学信号，解析得到第三待测试样中的第三粒子信息，第三粒子信息至少包括嗜碱性粒细胞信息；根据光接收部件反馈的第四光学信号，解析得到第四待测试样中的第四粒子信息，第四粒子信息包括网织红细胞信息。当第二反应装置2116包括第三反应池、第四反应池和第五反应池时，控制器2120分时复用于：根据光接收部件反馈的信息，解析得到样本的原始细胞计数信息、白细胞分类计数信息和网织红细胞计数信息。
作为一种实施方式，第二反应装置2116至少包括第三反应池，控制器2120还被配置为：在第一粒子信息的原始细胞信息中，扣减第二粒子信息中的异常淋巴细胞数据，得到原始细胞的测量数据。异常淋巴细胞会影响原始敏感区域粒子的识别，通过DIFF检测通道测得异常淋巴细胞的数据，并在原始细胞通道测得的数据扣减DIFF检测通道测得的异常淋巴细胞数据，从而得到更准确的原始细胞数据。
作为一种实施方式，第二反应装置2116至少包括第四反应池，控制器2120还被配置为：在第一粒子信息的原始细胞信息中，扣减第三粒子信息中的嗜碱性粒细胞数据，得到原始细胞的测量数据。嗜碱性粒细胞会影响原始敏感区域粒子的识别，通过WNB检测通道测得嗜碱性粒细胞的数据，并在原始细胞通道测得的数据扣减WNB检测通道测得的嗜碱性粒细胞数据，从而得到更准确的原始细胞数据。
作为一种实施方式，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，控制器2120被配置为控制样本分配装置2111按顺序执行如下动作：从样本容器2200中吸取样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至第一反应装置2113，将吸取的部分样本分配至第二反应装置2116，其中，样本分配装置2111从样本容器2200一次性吸取的样本量大于或等于分配至第一反应装置2113的样本量与分配至第二反应装置2116的样本量之和。本实施方案中，当一个样本容器2200中的样本既要做原始细胞检测项目，又要做血常规检测项目时，样本分配装置2111从样本容器2200中一次吸取原始细胞检测项目和血常规检测项目的样本量，然后吸取经 荧光标记的抗体试剂，将抗体试剂与原始细胞检测项目的样本分配至原始细胞反应通道，再将血常规检测项目的样本分配至血常规反应通道。由于原始细胞检测的反应时间比血常规检测的反应时间长，所以先分配原始细胞反应通道的样本，再分配血常规检测通道的样本，利于缩短同一个样本的检测结果输出时间。
当然，具体应用中，样本分析仪2100的分样顺序不限于上述方式。例如，作为上述分样方式的第一种替代实施方案，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，控制器2120被配置为控制样本分配装置2111按顺序执行如下动作：从样本容器2200中吸取样本，将吸取的部分样本分配至第二反应装置2116，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至第一反应装置2113，其中，样本分配装置2111从样本容器2200一次性吸取的样本量大于或等于分配至第一反应装置2113的样本量与分配至第二反应装置2116的样本量之和。该替代的实施方案中，一次吸样，先对血常规检测项目进行分样，再吸取抗体试剂，然后对原始细胞检测项目进行分抗体试剂和分样。
或者，作为上述分样方式的第二种替代实施方案，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，控制器2120被配置为控制样本分配装置2111按顺序执行如下动作：从样本容器2200中吸取第一份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第一份样本分配至第一反应装置2113，从样本容器2200中吸取第二份样本，将吸取的至少部分第二份样本分配至第二反应装置2116。该替代的实施方案中，原始细胞检测和血常规检测分开吸样，先对原始细胞检测项目进行吸样、吸抗体试剂、分抗体试剂和分样，然后再对血常规检测项目进行吸样、分样。
或者，作为上述分样方式的第三种替代实施方案，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，控制器2120被配置为控制样本分配装置2111按顺序执行如下动作：从样本容器2200中吸取第一份样本，将吸取的至少部分第一份样本分配至第二反应装置2116，从样本容器2200中吸取第二份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第二份样本分配至第一反应装置2113。该替代的实施方案中，原始细胞检测和血常规检测分开吸样，先对血常规检测项目进行吸样、分样，然后再对原始细胞检测项目进行吸样、吸抗体试剂、分抗体试剂和分样。
或者，作为上述分样方式的第四种替代实施方案，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，控制器2120被配置为控制样本分配装置2111按顺序执行如下动作：从样本容器2200中吸取样本，将吸取的部分样本分配至第二反应装置2116，将吸取的部分样本分配至第一反应装置2113，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂分配至第一反应装置2113，其中，样本分配装置2111从样本容器2200一次性吸取的样本量大于或等于分配至第一反应装置2113的样本量与分配至第二反应装置2116的样本量之和。该替代的实施方案中，样本和抗体试剂分开分配，且一次吸样，先对血常规检测项目进行分样，再对原始细胞检测项目进行分样，再吸取抗体试剂和分配抗体试剂。当然，在一次吸样后，也可以先原始细胞检测项目进行分样，再对血常规检测项目进行分样，然后再吸取抗体试剂和分配抗体试剂。
作为一种实施方式，当样本分配装置2111从样本容器2200一次性吸取的样本至少用于分别分配至第一反应装置2113和第二反应装置2116时，样本分配装置2111从样本容器2200一次性吸取的样本量小于或等于200μL。本实施方案中，原始细胞和血常规检测的样本用量小于或等于200μL，极大程度地减小样本用量。
作为一种实施方式，样本分析仪2100还包括血红蛋白检测装置2117，样本分配装置2111还用于将吸取的至少部分样本分配至血红蛋白检测装置2117，血红蛋白检测装置2117用于对至少由样本与第四类试剂制成的第五待测试样进行血红蛋白参数测定，控制器2120还被配置为：根据血红蛋白检测装置2117反馈的信息，解析得到第五待测 试样中的血红蛋白信息。血红蛋白检测属于血常规检测项目的一部分，当然，血常规检测中也不一定要检测血红蛋白。本实施方案提供的样本分析仪2100，还可以进行血红蛋白检测项目的测定，利于扩大样本分析仪2100的适用范围。
作为一种实施方式，第四类试剂包括稀释液和第四溶血试剂，即：第五待测试样由稀释液、第四溶血试剂制成。稀释液用于稀释样本。第四溶血试剂例如为能够溶解血液样本中的红细胞、释放红细胞中的血红蛋白并将血红蛋白转化为高铁血红蛋白的溶血剂。当然，具体应用中，稀释液和第四溶血试剂也可以合为一种试剂。
作为一种实施方式，样本分析仪2100还包括阻抗检测装置2118，阻抗检测装置2118用于对至少由样本与第五类试剂制成的第六待测试样进行测定，控制器2120还被配置为：根据阻抗检测装置2118反馈的信息，解析得到第六待测试样中的血小板信息和/或红细胞信息。血小板和红细胞属于血常规检测项目的一部分，当然，血常规检测中也不一定要检测血小板和/或红细胞。本实施方案提供的样本分析仪2100，还可以进行血小板和/或红细胞检测项目的测定，利于扩大样本分析仪2100的适用范围。
作为一种实施方式，第五类试剂为稀释液。
作为一种实施方式，血红蛋白检测装置2117包括第六反应池和血红蛋白检测组件，第六反应池至少用于为样本与第四类试剂提供反应场所以制备形成第五待测试样。第六反应池为血红蛋白的反应通道，即HGB反应通道。血红蛋白检测组件用于对第六反应池中的第五待测试样进行血红蛋白检测。
作为一种实施方式，阻抗检测装置2118包括检测室和阻抗检测组件，样本分析仪2100还包括第二试样输送组件，第六反应池还用于为样本与稀释液提供反应场所以制备形成第六待测试样，第二试样输送组件连接于第六反应池与检测室之间以用于在控制器2120的控制下将第六待测试样从第六反应池输送至检测室。本实施方案中，第六反应池先用于制备阻抗检测用的第六待测试样，通过第二试样输送组件将部分第六待测试样输送至检测室进行阻抗检测后，再在采用第六反应池中剩余的至少部分第六待测试样进行制备第五待测试样，相当于第六反应池复用于制备阻抗检测用的待测试样和血红蛋白检测用的待测试样。当然，具体应用中，也可以为阻抗检测设置单独的反应池进行制备阻抗检测用的待测试样。
作为一种实施方式，样本分配装置2111包括样本针21111、样本吸排驱动部件和针运动驱动部件；样本针21111用于吸取和排放样本。样本吸排驱动部件用于为样本吸、排样本提供驱动力。针运动驱动部件用于驱动样本针21111进行二维或三维空间运动，以使样本针21111分别运动至不同的工位，例如分别运动至待机位、吸样位、加样位、吸抗体试剂位、清洗位等。
作为一种实施方式，样本吸排驱动部件为注射器。
作为一种实施方式，样本分析仪2100对一个样本容器2200内样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果。如果一个患者，既需要做原始细胞检测项目，又要做血常规检测项目，则从患者采集的血液样本可以共用一个样本容器2200，即两种类型的检测项目可以共用一个样本容器2200装载样本，这样，一方面利于减少样本容器2200的消耗，另一方面利于减少所要采集的样本量。当然，具体应用中，作为替代的实施方案，原始细胞检测项目检测用的样本和血常规检测项目检测用的样本也可以分开装在两个不同的样本容器2200内。
作为一种实施方式，样本分析仪2100对样本分配装置2111从样本容器2200中一次吸取的样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果。本实施方案中，一次吸样至少输出两种检测结果，可以利于减小样本分配装置2111的吸样动作次数，从而利于提高样本分析仪2100的检测效率。当然，具体应用中，作为替代的实施方案，对于两种以上的检测项目，也可以分开独立吸样，即对一个样本容 器2200中样本的吸样次数也可以两次以上。
作为一种实施方式，样本分析仪2100还包括识别装置2119，识别装置2119用于对样本容器2200进行识别。控制器2120还被配置为：根据识别装置2119的反馈信息，至少获取样本容器2200内样本的待测检测项目信息。
作为一种实施方式，控制器2120还被配置为：根据待测检测项目信息，控制样本分析仪2100对样本容器2200内的样本进行原始细胞检测和/或血常规检测。本实施方案中，样本分析仪2100根据识别装置2119的反馈信息，自动获取样本的检测项目信息，并自动控制分析仪执行主体2110执行相对应的检测项目，自动化程度高。
作为一种实施方式，样本分析仪2100还包括人机交互装置2140，人机交互装置2140配置有原始细胞检测模式和血常规检测模式，控制器2120还被配置为：根据操作人员在人机交互装置2140选择的检测模式，控制样本分析仪2100对样本容器2200内的样本进行原始细胞检测和/或血常规检测。本实施方案中，操作人员可以通过人机交互装置2140，手动启动样本的检测模式。
作为一种实施方式，人机交互装置2140包括显示屏、键盘、鼠标中的至少一种。
作为一种实施方式，识别装置2119用于识别样本容器2200的标识码，标识码包括二维码、条形码、无线射频(RFID)中的至少一种。控制器2120根据识别装置2119识别的标识码，获取样本容器2200中样本的检测项目信息。
当然，具体应用中，作为替代的实施方案，识别装置2119除了用于识别样本容器2200的标识码外，还可以用于识别样本容器2200的外观特征。样本容器2200的外观特征包括样本容器2200的容器主体形状、容器主体大小、样本量、是否有帽体、帽体的颜色、帽体的形状、帽体的大小中的至少一种。控制器2120根据识别装置2119的反馈信息，除了获取样本的检测项目信息外，还至少获取如下信息中的一种：样本容器200中的样本量，样本容器200的类型，样本编号信息。
作为一种实施方式，识别装置2119包括视觉相机、条形码扫描仪、二维码扫描仪、无线射频读码仪中的至少一种。
以上仅示例性地说明了本公开的细胞分析仪的结构和构成部件。本领域技术人员能够据此得到各种修改、变形或替代的方案均包含在本公开的范围内。以下通过进一步的实施例说明本公开的方法。
实施例1
试剂A：DIFF荧光染料(M-68P FD染色液)20μL
试剂B：DIFF通道溶血剂(M-68P LD溶血剂)1mL
试剂C：CD45-FITC(Biolegend，200μg/mL，pH＝7.2，磷酸盐缓冲液，0.2％牛血清白蛋白，0.09％叠氮钠)1μL
血细胞分析仪(Mindray BC-6800Plus，激发波长450nm)通过采样针吸取20微升待测血样(取自受试者的末梢血，含未成熟粒细胞(IG))送入混合室。试剂A～C按照上述量加入到混合室，与血样混合后，42℃条件下孵育60秒后，制备成待测试样并进行检测收集上述待测试样中的粒子产生前向散射光、侧向散射光和荧光信号。
设备DIFF通道测得该样本白细胞含量为3.4×109/L，在CD45FL/SSC的维度通过对划分的BLAST区域内的散点数进行计数，得出BLAST％为0％，因此对FL/SSC无需校正。根据FL/SSC散点图(图13)，获得白细胞四分类结果为Neu％、Lym％、Mon％、Eos％和IG％分别为64.2％、7.1％、9.2％，4.4％和21.1％。上述比例可进一步通过计算转化为样本中各粒子的含量。
另外取相同血液样本通过经典人工镜检计数的方法获得的Neu％、Lym％、Mon％、Eos％和IG％分别为62.3％、6.61％、7.4％、4％％和18.1％。
从上述结果可以看出，上述方法对白细胞进行划分与人工镜检中的比例具有较好的相关性。
实施例2
试剂A：DIFF荧光染料(M-68P FD染色液)20μL
试剂B：DIFF通道溶血剂(M-68P LD溶血剂)1mL
试剂C：CD45-FITC(Biolegend，200μg/mL，pH＝7.2，磷酸盐缓冲液，0.2％牛血清白蛋白，0.09％叠氮钠)1μL
按照与实施例1相同的方法并使用相同的设备对待测血样(取自受试者的末梢血，含有原始细胞)进行处理和检测。
设备DIFF通道测得该样本的白细胞(包括原始细胞)含量为12.6×109/L。在CD45FL/SSC的维度通过对划分的BLAST区域内的散点数进行计数，得出BLAST％为12％，进而通过计算得到样本中BLAST含量为1.5×109/L。根据CD45FL/SSC的维度划分为BLAST的粒子，确定其在FL/SSC维度的分布情况(图14虚线框)，扣除掉FL/SSC维度中上述区域BLAST粒子后，获得校正的Lym和Mon区域中粒子数，可以获得校正后的分类结果：Neu％、Lym％、Mon％和Eos％分别为52.9％、33.7％、1.1％和0.2％。
另外取相同血液样本通过经典人工镜检计数的方法获得的BLAST％、Neu％、Lym％、Mon％、Eos％和IG％分别为10.4％、53.2％、31.4％、1.2％和0.2％。从结果可以看出，上述方法对原始细胞以及白细胞进行划分与人工镜检中的比例具有较好的相关性。
实施例3
试剂A：DIFF荧光染料(Mindray，M-68P FD染色液)20μL
试剂B：DIFF通道溶血剂(Mindray，M-68P LD溶血剂)1mL
试剂C：CD45-FITC(Biolegend，200μg/mL，ph＝7.2，磷酸盐缓冲液，0.2％牛血清白蛋白，0.09％叠氮钠)1μL
试剂D：CD45通道溶血剂(Mindray，二甘醇、甲醛、缓冲液)1mL
试剂E：WNB荧光染料(M-68P FN染色液)20μL
试剂F：WNB通道溶血剂(M-68P LN溶血剂)1mL
血细胞分析仪(Mindray BC-6800Plus，激发波长450nm)通过采样针吸取60微升待测血样(与实施例2相同的血样)，分成三份待测子血样分别进入不同的混合室。试剂A和B按照上述用量加入到DIFF通道混合室，与血样混合后42℃条件下孵育30秒后，制备成第二待测试样；试剂E和F按照上述用量加入到WNB通道混合室，与血样混合后42℃条件下孵育30秒后，制备成第三待测试样；试剂C和D按上述用量加入到CD45通道混合室，与血样混合后42℃条件下孵育45秒后，制备成第一待测试样。对三个待测试样依次进行检测并分别收集上述各通道试样中的粒子产生前向散射光、侧向散射光和荧光信号。
设备DIFF通道测得该样本白细胞含量为12.6×109/L。在WNB通道的FL/SSC维度得到Lym％为33.7％(参见图15中的A图)。在CD45FL/SSC的维度通过对划分的BLAST区域内的散点数进行计数，得出BLAST％为12％，进而通过计算得到样本中原始细胞含量为1.5×109/L。根据CD45FL/SSC维度得到BLAST％(12％)，WNB通道FL/SSC维度得到Lym％(33.7％)，结合上述两个结果对应在DIFF通道进行比例扣除得到校正后的Neu％、Mon％和Eos％分别为52.9％、1.1％和0.2％。
与实施例2中通过经典人工镜检计数的方法获得的BLAST％、Neu％、Lym％、Mon％、Eos％和IG％分别为10.4％、53.2％、31.4％、1.2％和0.2％的结果可以看出，上述方法对白细胞进行划分与人工镜检中的比例具有较好的相关性。
实施例4
试剂C：CD45-FITC(Biolegend，200μg/mL,ph＝7.2,磷酸盐缓冲液，0.2％牛血清白蛋白，0.09％叠氮钠)1μL
试剂D：CD45通道溶血剂(Mindray，二甘醇、甲醛、缓冲液)1mL
血细胞分析仪(Mindray BC-6800Plus，激发波长450nm)通过采样针吸取20微升待测血样(取自患者的末梢血)，送入入混合室，试剂C和D按照上述用量加入到混合室，与血样混合后42℃条件下孵育60秒后，制备成待测试样并进行检测，收集上述试样中的粒子产生前向散射光、侧向散射光和荧光信号。
设备测得该样本中白细胞含量为11.9×109/L。在CD45FL/SSC的维度(参见图16)对划分的BLAST、Neu、Lym、Mon和Eos各自区域内的散点数进行计数，得出BLAST％、Neu％、Lym％、Mon％和Eos％分别为11.9％、52.9％、33.7％、1.1％和0.2％。
另外取相同血液样本通过经典人工镜检计数的方法获得的BLAST％、Neu％、Lym％、Mon％、Eos％和IG％分别为10.4％、53.2％、31.4％、1.2％和0.2％，从上述结果可以看出，上述方法对原始细胞和白细胞进行划分与人工镜检中的比例具有较好的相关性。
以上所述仅为本发明的优选实施方式，并非因此限制本发明的专利范围，凡是在本发明的发明构思下，利用本发明说明书及附图内容所作的等效结构变换，或直接/间接运用在其他相关的技术领域均包括在本发明的专利保护范围内。

Claims (48)

  1. 一种血液分析仪，包括吸样装置、试剂供应装置、混合室、光源、光学流动室、光学检测器、处理器和编程有计算机应用程序的非暂时性计算机可读存储介质，其中，
    所述吸样装置用于吸取样本，并将所述吸取样本的至少部分作为第一子样本输送到所述混合室；
    所述试剂供应装置用于提供试剂并输送到所述混合室，所述试剂包括第一类试剂；
    所述混合室用于将所述第一子样本与所述第一类试剂混合以形成第一待测试样；其中所述第一类试剂包括第一试剂和第二试剂，所述第一试剂包括第一溶血剂，所述第二试剂包括经荧光标记的抗体，所述抗体能够与原始细胞的表面抗原相结合；
    所述光源用于将光束对准与所述混合室相连通的所述光学流动室的检测孔；
    所述光学检测器用于检测通过所述检测孔的所述第一待测试样的光散射信号和荧光信号，所述光散射信号包括来自第一待测试样的第一侧向光散射强度信号，所述荧光信号包括来自所述经荧光标记的抗体的抗体荧光强度信号；
    所述处理器与所述光学检测器可操作地连接，当所述非暂时性计算机可读存储介质中的计算机应用程序被所述处理器执行时，控制所述第一溶血剂与所述第一子样本在所述混合室中的混合时间不超过2分钟，且所述处理器根据所述第一侧向光散射强度信号和所述抗体荧光强度信号获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括原始细胞信息。
  2. 根据权利要求1所述的血液分析仪，其中，
    所述第一类试剂还包括第三试剂，所述第三试剂包括第一荧光染料；
    所述光散射信号还包括来自所述第一荧光染料的第一荧光信号；和
    当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一侧向光散射强度信号和所述第一荧光强度信号，获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息。
  3. 根据权利要求2所述的血液分析仪，其中，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本中不含原始细胞时，根据所述第一子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息，或者
    所述处理器根据所获得的原始细胞信息，当所述原始细胞信息显示所述样本中含有原始细胞时，从所述第一子样本的粒子信息中扣除所述原始细胞的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
  4. 根据权利要求1所述的血液分析仪，其中，
    所述吸样装置还将所述吸取样本的至少部分作为第二子样本和第三子样本依次输送到所述混合室；
    所述试剂还包括第二类试剂，所述第二类试剂包括第三试剂、第四试剂、第五试剂和第六试剂；
    所述混合室进一步用于将所述第二和第三子样本分别进行混合，其中第二子样本与第三试剂和第四试剂混合以获得第二待测试样，第三子样本与第五试剂和第六试剂混合以获得第三待测试样；其中，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同， 所述第二荧光染料不同于所述第一荧光染料；
    所述光学检测器进一步用于检测分别通过所述检测孔的所述第二和第三待测试样的光散射信号和荧光信号；所述光散射信号包括来自第二待测试样的第二侧向光散射强度信号、以及来自第三待测试样的第三侧向光散射强度信号；所述荧光信号包括来自第二待测试样的第一荧光染料的第二荧光强度信号和来自第三待测试样的第二荧光染料的第三荧光强度信号；
    当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第二待测试样的所述第二侧向光散射强度信号和所述第二荧光强度信号获得所述第二子样本中的粒子信息，和根据所述第三待测试样的所述第三侧向光散射强度信号和所述第三荧光强度信号获得所述第三子样本中的粒子信息；其中所述第二子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息，所述第三子样本中的粒子信息包括淋巴细胞信息、嗜碱性粒细胞信息和有核红细胞信息；
    其中，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本不含原始细胞时，根据所述第二子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息；或者
    当所述原始细胞信息显示所述样本含有原始细胞时，根据所述第二子样本中的粒子信息，结合所述原始细胞信息和所述第三子样本中的粒子信息中的淋巴细胞信息，消除所述原始细胞信息对所述第二子样本中的粒子信息的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息。
  5. 根据权利要求1～4中任一项所述的血液分析仪，其中，当所述计算机应用程序被所述处理器执行时，使所述处理器对原始细胞进行计数。
  6. 根据权利要求1～4中任一项所述的血液分析仪，其中，当所述计算机应用程序被所述处理器执行时，所述处理器进一步，当所述原始细胞信息显示所述样本含有原始细胞时，报警所述样本中存在原始细胞，或报警所述样本中存在原始细胞并输出原始细胞计数值。
  7. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一前向光散射强度信号和所述抗体荧光强度信号确定所述原始细胞为淋系原始细胞和/或髓系原始细胞。
  8. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，当所述计算机应用程序被所述处理器执行时，所述处理器进一步根据所述第一前向光散射强度信号和所述抗体荧光强度信号报警所述样本中存在淋系原始细胞和/或髓系原始细胞。
  9. 根据权利要求1～4中任一项所述的血液分析仪，其中，当所述计算机应用程序被所述处理器执行时，所述处理器根据所述第一侧向光散射强度信号和抗体荧光强度信号进一步获得所述样本中的白细胞信息，所述白细胞信息包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的信息。
  10. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述第一溶血剂与所述样本的作用时间为30～80秒，优选为50～60秒。
  11. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述第一溶血剂能够使红细胞完全破碎，同时白细胞维持完整的细胞形态。
  12. 根据权利要求11所述的血液分析仪，其中，所述第一溶血剂包括选自季铵盐类阳离子表面活性剂、烷醚乙氧类非离子表活剂、烷醇聚氧乙烯醚类表面活性剂、烷基 糖苷、三萜皂苷、甾族皂苷和二甘醇中的至少一种，优选地，所述第一溶血剂包括溶解在缓冲液中的二甘醇和甲醛。
  13. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述抗体为CD45。
  14. 根据权利要求1～4中任一项所述的血液分析仪，其中，所述血液分析仪还包括血红蛋白检测单元和/或阻抗计数检测单元。
  15. 一种样本分析方法，所述方法包括：
    接收操作指令，对样本进行原始细胞检测；
    其中，所述原始细胞检测包括：
    吸取样本，包括吸取所述样本的至少部分作为第一子样本；
    处理样本，包括将所述第一子样本与第一类试剂混合以获得第一待测试样；其中所述第一类试剂包括第一试剂和第二试剂，所述第一试剂包括第一溶血剂，所述第一溶血剂与所述第一子样本的混合时间不超过2分钟，所述第二试剂包括荧光标记的抗体，所述抗体能够与原始细胞的表面抗原相结合；
    检测待测试样，包括使所述第一待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源对所述第一待测试样中的粒子进行照射，以测得第一待测试样的光散射信号和荧光信号，所述光散射信号包括来自第一待测试样的第一侧向光散射强度信号，所述荧光信号包括来自所述荧光标记的抗体的抗体荧光强度信号；和
    获取粒子信息，包括根据所述第一侧向光散射强度信号和所述抗体荧光强度信号获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括原始细胞信息。
  16. 根据权利要求15所述的样本分析方法，其中，所述第一类试剂还包括第三试剂，所述第三试剂包括第一荧光染料，且所述光散射信号还包括来自第一待测试样中第一荧光染料的第一荧光信号；
    所述原始细胞检测进一步包括：根据所述第一侧向光散射强度信号和所述第一荧光强度信号，获得所述第一子样本中的粒子信息，所述第一子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息。
  17. 根据权利要求16所述的样本分析方法，其中，所述原始细胞检测进一步包括：
    当所述原始细胞信息显示所述样本中不含原始细胞时，根据所述第一子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息；或者
    当所述原始细胞信息显示所述样本中含有原始细胞时，从所述第一子样本中的粒子信息中扣除所述原始细胞的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
  18. 根据权利要求15所述的样本分析方法，其中，在所述原始细胞检测中，
    所述吸取样本进一步包括吸取所述样本的至少部分作为第二子样本和第三子样本；
    所述处理样本进一步包括用第二类试剂处理所述第二和第三子样本，其中所述第二类试剂包括第三试剂、第四试剂、第五试剂和第六试剂，将所述第二子样本与第三试剂和第四试剂混合以获得第二待测试样，和将所述第三子样本与第五试剂和第六试剂混合以获得第三待测试样；其中，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同，以及所述第二荧光染料不同于所述第一荧光染料；
    所述检测待测试样进一步包括分别使所述第二和第三待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源分别对所述第二和第三待测试样中的粒子进行照射，以测得第二和第三待测试样的光散射信号和荧光信号；其中，所述光散射信号包括来自第二待测试样的第二侧向光散射强度信号、以及来自第三待测 试样的第三侧向光散射强度信号；所述荧光信号包括来自第二待测试样中第一荧光染料的第二荧光强度信号和来自第三待测试样中第二荧光染料的第三荧光强度信号；
    所述获取粒子信息进一步包括根据所述第二待测试样的所述第二侧向光散射强度信号和所述第二荧光强度信号获得所述第二子样本中的粒子信息，和根据所述第三待测试样的所述第三侧向光散射强度信号和所述第三荧光强度信号获得所述第三子样本中的粒子信息；其中所述第二子样本中的粒子信息包括中性粒细胞信息、单核细胞信息、淋巴细胞信息和嗜酸性粒细胞信息，所述第三子样本中的粒子信息包括淋巴细胞信息、嗜碱性粒细胞信息和有核红细胞信息；以及，
    所述原始细胞检测进一步包括：
    当所述原始细胞信息显示所述样本不含原始细胞时，根据所述第二子样本中的粒子信息，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息；或者
    当所述原始细胞信息显示所述样本含有原始细胞时，根据所述第二子样本中的粒子信息，结合所述原始细胞信息和所述第三子样本中的粒子信息中的淋巴细胞信息，消除所述原始细胞信息对所述第二子样本中的粒子信息的影响，获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和异型淋巴细胞信息。
  19. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述方法还包括对样本进行常规检测；
    其中所述常规检测包括：
    吸取所述样本的至少部分作为第一初始子样本；
    处理所述第一初始子样本，包括将所述第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样，其中所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同；
    使所述第一初始待测试样中的粒子逐个通过光学检测装置的检测区并且利用所述光学检测装置的光源对所述第一初始待测试样中的粒子进行照射，以获取所述第一初始待测试样中粒子的光散射信号和荧光信号，所述光散射信号包括来自第一初始待测试样的第一初始侧向光散射强度信号；所述荧光信号包括来自第一初始待测试样中第一荧光染料的第一初始荧光信号；
    根据所述第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括白细胞信息；和
    当所述白细胞信息异常时，发出原始细胞报警，并可选地发出所述操作指令。
  20. 根据权利要求15所述的样本分析方法，其中，所述方法还包括对样本进行常规检测；
    其中所述常规检测包括：
    分别吸取所述样本的至少部分作为第一初始子样本和第二初始子样本；
    分别处理所述第一和第二初始子样本，包括将所述第一初始子样本与第三试剂和第四试剂混合以获得第一初始待测试样，和将所述第二初始子样本与第五试剂和第六试剂混合以获得第二初始待测试样，所述第三试剂包括第一荧光染料，所述第四试剂包括第二溶血剂，所述第五试剂包括第二荧光染料，所述第六试剂包括第三溶血剂，且所述第二溶血剂与所述第一溶血剂相同或不同，所述第三溶血剂与所述第一和第二溶血剂均不同，以及所述第二荧光染料不同于所述第一荧光染料；
    分别使所述第一和第二初始待测试样中的粒子逐个通过光学检测装置的检测区，并且利用所述光学检测装置的光源分别对所述第一和第二初始待测试样中的粒子进行照射，以获取所述第一和第二初始待测试样中粒子的光散射信号和荧光信号，所述光散射 信号包括来自第一初始待测试样的第一初始侧向光散射强度信号和来自第二初始待测试样的第二初始侧向光散射强度信号；所述荧光信号包括来自第一初始待测试样中第一荧光染料的第一初始荧光信号和来自第二初始待测试样中第二荧光染料的第二初始荧光信号；
    根据所述第一初始侧向光散射强度信号和第一初始荧光强度信号获得所述第一初始待测试样的粒子信息，所述粒子信息至少包括初始白细胞信息，和根据所述第二初始侧向光散射强度信号和第二初始荧光强度信号获得所述第二初始待测试样的粒子信息，所述粒子信息至少包括初始淋巴信息；和
    当所述初始白细胞信息异常时，发出原始细胞报警，并可选地发出所述操作指令。
  21. 根据权利要求20所述的样本分析方法，其中，所述对样本进行原始细胞检测进一步包括：
    结合所述原始细胞信息和所述初始淋巴细胞信息，消除所述原始细胞对所述初始白细胞信息的影响，并获得中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞的分类计数，以及可选地获得未成熟粒细胞信息和/或异型淋巴细胞信息。
  22. 根据权利要求19～21中任一项所述的样本分析方法，其中，所述常规检测在所述原始检测之前进行，进一步地可以根据常规检测的结果发出所述操作指令。
  23. 根据权利要求22所述的样本分析方法，其中，所述操作指令包括用户输入的指令或当发出原始细胞报警时自动触发的指令。
  24. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述方法进一步包括：对原始细胞进行计数。
  25. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述方法进一步包括：当所述原始细胞信息显示所述样本含有原始细胞时，报警所述样本中存在原始细胞，或报警所述样本中存在原始细胞并输出原始细胞计数值。
  26. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，所述方法进一步包括：
    根据所述第一前向光散射强度信号和所述抗体荧光强度信号确定所述原始细胞为淋系原始细胞和/或髓系原始细胞。
  27. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述光散射信号还包括来自第一待测试样的第一前向散射光信号，所述方法进一步包括：
    根据所述第一前向光散射强度信号和所述抗体荧光强度信号报警所述样本中存在淋系原始细胞和/或髓系原始细胞。
  28. 根据权利要求15～18中任一项所述的样本分析方法，其中，所述方法进一步包括：根据所述第一侧向光散射强度信号和抗体荧光强度信号进一步获得所述样本中的白细胞信息，所述白细胞信息包括淋巴细胞、单核细胞、中性粒细胞和嗜酸性粒细胞的信息。
  29. 根据权利要求15～19中任一项所述的样本分析方法，其中，所述第一溶血剂与所述样本的作用时间为30～80秒，优选为50～60秒。
  30. 根据权利要求15～19中任一项所述的样本分析方法，其中，所述第一溶血剂能够使红细胞完全破碎，同时白细胞维持完整的细胞形态。
  31. 根据权利要求15～19中任一项所述的样本分析方法，其中，所述第一溶血剂为选自季铵盐类阳离子表面活性剂、烷醚乙氧类非离子表活剂、烷醇聚氧乙烯醚类表面活性剂、烷基糖苷、三萜皂苷、甾族皂苷和二甘醇中的至少一种，优选地，所述第一溶血剂包括溶解在缓冲液中的二甘醇和甲醛。
  32. 根据权利要求15～19中任一项所述的样本分析方法，其中，所述抗体为CD45。
  33. 一种样本分析仪，包括样本分配装置、光学检测装置、第一反应装置、第二反 应装置、第一试样输送组件和控制器；
    所述样本分配装置用于从样本容器中吸取样本，并将吸取的至少部分样本分配至第一反应装置和/或所述第二反应装置；
    所述第一反应装置至少用于为样本与抗体试剂、第一溶血试剂提供反应场所以制备形成第一待测试样，其中，所述抗体试剂中的抗体能够与样本中原始细胞的表面抗原相结合；
    所述第二反应装置至少用于为样本与第一类试剂提供反应场所以制备形成第二待测试样；
    所述第一试样输送组件用于将所述第一待测试样从所述第一反应装置输送至所述光学检测组件，且用于将所述第二待测试样从所述第二反应装置输送至所述光学检测组件；
    所述光学检测组件包括流动室、光发射部件和光接收部件，所述流动室用于供所述第一待测试样或所述第二待测试样在鞘液的裹挟下通过，所述光发射部件用于朝向所述流动室中的第一待测试样或所述第二待测试样发射光线，所述光接收部件用于接收所述光发射部件发射的光线经所述第一待测试样产生的第一光学信号以及用于接收所述光发射部件发射的光线经所述第二待测试样产生的第二光学信号；
    所述控制器被配置为：根据所述光接收部件反馈的所述第一光学信号，解析得到所述第一待测试样中的第一粒子信息，所述第一粒子信息包括原始细胞信息；
    根据所述光接收部件反馈的所述第二光学信号，解析得到所述第二待测试样中的第二粒子信息，所述第二粒子信息包括中性粒细胞信息、嗜酸性粒细胞信息、单核细胞信息和淋巴细胞信息。
  34. 如权利要求33所述的样本分析仪，其中，所述样本分析仪还包括外壳，所述光学检测装置、所述第一反应装置和所述第二反应装置都位于所述外壳内。
  35. 如权利要求33所述的样本分析仪，其中，所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的样本量小于或等于60uL；且/或，
    所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的样本量在50uL±10uL之间。
  36. 如权利要求33所述的样本分析仪，其中，所述控制器还被配置为：在单个原始细胞检测项目的检测过程中，控制样本与抗体试剂在所述第一反应装置的反应时间小于或等于90s，优选为小于或等于60s。
  37. 如权利要求33所述的样本分析仪，其中，所述抗体试剂为CD45试剂。
  38. 如权利要求33所述的样本分析仪，其中，所述样本分配装置还用于从第一试剂容器中吸取所述抗体试剂，并将吸取的所述抗体试剂分配至所述第一反应装置；或者，
    所述样本分析仪还包括第一试剂分配装置，所述第一试剂分配装置用于从第一试剂容器中吸取所述抗体试剂，并将吸取的所述抗体试剂分配至所述第一反应装置。
  39. 如权利要求33所述的样本分析仪，其中，所述抗体试剂中包含经荧光标记、且能够与原始细胞表面抗原相结合的抗体，所述第一光学信号包括由所述光发射部件发射的光线照射至所述第一待测试样产生的第一侧向散射光信号和由所述光发射部件发射的光线照射至所述第一待测试样经所述荧光标记产生的第一荧光信号；或者，
    所述抗体试剂中包含未经荧光标记、但能够与原始细胞表面抗原相结合的抗体，所述第二反应池用于为样本与所述抗体试剂、第一荧光试剂提供反应场所以制备形成所述第一反应液，所述第一光学信号包括由所述光发射部件发射的光线照射至所述第一待测试样产生的第一侧向散射光信号和由所述光发射部件发射的光线照射至所述第一待测试样经所述第一荧光试剂产生的第一荧光信号。
  40. 如权利要求39所述的样本分析仪，其中，所述第一反应装置用于为所述第一反应液与所述第一溶血试剂、第二荧光试剂提供反应场所以制备形成所述第一待测试样；
    所述第一光学信号还包括由所述光发射部件发射的光线照射至所述第一待测试样经所述第二荧光试剂产生的第二荧光信号；
    所述控制器根据所述光接收部件反馈的所述第一光学信号，解析得到所述第一待测试样中的第一粒子信息：根据所述第一侧向散射光信号和所述第一荧光信号解析得到第一原始细胞信息和第一血影区域信息，根据所述第二荧光信号解析得到第二血影区域信息；
    将所述第一原始细胞信息和所述第一血影区域信息，扣减所述第二血影区域信息，得到第一待测试样的原始细胞信息。
  41. 如权利要求33至40任一项所述的样本分析仪，其中，所述第二反应装置包括第三反应池，且包括第四反应池和第五反应池中的至少一者，所述第三反应池用于为样本与所述第一类试剂提供反应场所以制备形成所述第二待测试样，所述第四反应池用于为样本与第二类试剂提供反应场所以制备形成第三待测试样，所述第五反应池用于为样本与第三类试剂提供反应场所以制备形成第四待测试样；
    所述第一试样输送组件还用于将所述第三待测试样或所述第四待测试样从所述第二反应装置输送至所述流动室；
    所述流动室还用于供第三待测试样或所述第四待测试样在鞘液的裹挟下通过，所述光发射部件还用于朝向所述流动室中的所述第三待测试样或所述第四待测试样发射光线，所述光接收部件还用于接收所述光发射部件发射的光线经所述第三待测试样产生的第三光学信号、或经所述第四待测试样产生的第四光学信号；
    所述控制器还被配置为根据所述光接收部件的反馈信息执行如下的至少一个解析动作：
    根据所述光接收部件反馈的所述第三光学信号，解析得到所述第三待测试样中的第三粒子信息，所述第三粒子信息至少包括嗜碱性粒细胞信息；
    根据所述光接收部件反馈的所述第四光学信号，解析得到所述第四待测试样中的第四粒子信息，所述第四粒子信息包括网织红细胞信息。
  42. 如权利要求41所述的样本分析仪，其中，当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至所述第一反应装置，将吸取的部分样本分配至所述第二反应装置，其中，所述样本分配装置从所述样本容器一次性吸取的样本量大于或等于分配至所述第一反应装置的样本量与分配至所述第二反应装置的样本量之和；或者，
    当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取样本，将吸取的部分样本分配至所述第二反应装置，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和部分样本分配至所述第一反应装置，其中，所述样本分配装置从所述样本容器一次性吸取的样本量大于或等于分配至所述第一反应装置的样本量与分配至所述第二反应装置的样本量之和；或者，
    当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取第一份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第一份样本分配至所述第一反应装置，从样本容器中吸取第二份样本，将吸取的至少部分第二份样本分配至所述第二反应装置；或者，
    当样本的检测项目包括原始细胞检测项目和血常规检测项目时，所述控制器被配置 为控制所述样本分配装置按顺序执行如下动作：从样本容器中吸取第一份样本，将吸取的至少部分第一份样本分配至所述第二反应装置，从样本容器中吸取第二份样本，从第一试剂容器中吸取抗体试剂，将吸取的抗体试剂和至少部分第二份样本分配至所述第一反应装置。
  43. 如权利要求42所述的样本分析仪，其中，当所述样本分配装置从所述样本容器一次性吸取的样本至少用于分别分配至所述第一反应装置和所述第二反应装置时，所述样本分配装置从所述样本容器一次性吸取的样本量小于或等于200uL。
  44. 如权利要求41所述的样本分析仪，其中，所述第二反应装置至少包括第三反应池，所述控制器还被配置为：在所述第一粒子信息的所述原始细胞信息中，扣减所述第二粒子信息中的异常淋巴细胞数据，得到原始细胞的测量数据；且/或，
    所述第二反应装置至少包括第四反应池，所述控制器还被配置为：在所述第一粒子信息的所述原始细胞信息中，扣减所述第三粒子信息中的嗜碱性粒细胞数据，得到原始细胞的测量数据。
  45. 如权利要求33至40任一项所述的样本分析仪，其中，所述控制器还被配置为：在单个原始细胞检测项目的样本分配过程中，控制所述样本分配装置分配至所述第一反应装置的抗体试剂的量小于5uL；且/或，
    所述控制器还被配置为：在单个原始细胞检测项目的抗体试剂分配过程中，控制分配至所述第一反应装置的抗体试剂的量在5uL±2uL之间。
  46. 如权利要求33至40任一项所述的样本分析仪，其中，所述样本分析仪还包括血红蛋白检测装置，所述样本分配装置还用于将吸取的至少部分样本分配至所述血红蛋白检测装置，所述血红蛋白检测装置用于对至少由样本与第四类试剂制成的第五待测试样进行血红蛋白参数测定，所述控制器还被配置为：根据所述血红蛋白检测装置反馈的信息，解析得到所述第五待测试样中的血红蛋白信息；且/或，
    所述样本分析仪还包括阻抗检测装置，所述阻抗检测装置用于对至少由样本与第五类试剂制成的第六待测试样进行测定，所述控制器还被配置为：根据所述阻抗检测装置反馈的信息，解析得到所述第六待测试样中的血小板信息和/或红细胞信息。
  47. 如权利要求33至40任一项所述的样本分析仪，其中，所述样本分析仪对一个样本容器内样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果；且/或，
    所述样本分析仪对所述样本分配装置从样本容器中一次吸取的样本的测定，至少输出如下两种检测结果：原始细胞检测结果，血常规检测结果。
  48. 如权利要求33至40任一项所述的样本分析仪，其中，所述样本分析仪还包括识别装置，所述识别装置用于对样本容器进行识别，所述控制器还被配置为：根据所述识别装置的反馈信息，至少获取样本容器内样本的待测检测项目信息，根据所述待测检测项目信息，控制所述样本分析仪对样本容器内的样本进行原始细胞检测和/或血常规检测；且/或，
    所述样本分析仪还包括人机交互装置，所述人机交互装置配置有原始细胞检测模式和血常规检测模式，所述控制器还被配置为：根据操作人员在所述人机交互装置选择的检测模式，控制所述样本分析仪对样本容器内的样本进行原始细胞检测和/或血常规检测。
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