Blood testing method and blood analysis system

Abstract

A blood testing method and a corresponding blood analysis system, the method comprising: treating a blood sample with a first reagent containing a hemolytic agent to obtain a test sample (S11), wherein the hemolytic agent lyses red blood cells in the blood sample into fragments; obtaining optical information of the test sample by an optical detection device (50, 200) (S12), wherein particles in the test sample are passed one by one through a detection zone of the optical detection device (50, 200) and the particles in the test sample are illuminated with a light source (101) of the optical detection device (50, 200) to obtain the optical information of the test sample, wherein the light source (101) is configured to emit light with a wavelength of less than 488 nm; and obtaining optical information of platelets (S13) and / or optical information of reticulocytes from at least two of the optical information, wherein the optical information of the platelets and / or the reticulocytes is obtained in a detection channel in which the red blood cells in the blood sample are lysed.

Description

Technical Field

[0001] This invention relates to blood testing, and particularly to an optical detection method for platelets and a blood analysis system thereof. Background Technology

[0002] Human blood contains various cells such as red blood cells, white blood cells, and platelets. Platelets are anucleate cells with a diameter of 2-3 micrometers. Normal human blood contains 150,000 to 350,000 platelets per microliter.

[0003] As is well known, the methods for measuring platelets in blood samples using a blood analyzer typically include impedance and optical methods.

[0004] Impedance method, based on the Coulter principle, involves passing particles from a diluted blood sample one by one through a small orifice. A constant current source is applied to both sides of the orifice. Each cell passing through the orifice causes a change in the electrical impedance of the liquid within the orifice, generating an electrical pulse. The detected electrical pulses are then plotted as a histogram for analysis. In normal blood, platelets are the smallest, white blood cells are the largest, and red blood cells are in between. The intensity of the detected pulse is related to the volume of the cells passing through the orifice, thus different cell types can be distinguished by volume division. However, testing certain special samples (such as those containing large platelets and small red blood cells) can affect the accuracy and precision of platelet detection. These special samples are usually from subjects with diseases, and deviations in the measured values ​​can adversely affect clinical diagnosis. Furthermore, in the impedance method, for low PLT samples, the boundary between the PLT histogram and the RBC histogram is often unclear, causing the algorithm to fail to accurately segment the PLT and RBC histograms, thus failing to obtain accurate PLT measurement results.

[0005] To overcome this limitation, an optical method for determining PLT has been proposed. This optical method is based on flow cytometry, in which diluted and stained samples are compressed by a sheath flow, causing cell particles in the sample to pass sequentially through an optical detection zone. Each cell is illuminated by an excitation light source, and the optical detection device obtains a forward scattering signal representing cell volume information and a fluorescence signal from particles stained with a fluorescent dye. This generates a two-dimensional scatter plot of the forward scattering and fluorescence signals, which is then used to classify and count RBCs and PLTs.

[0006] Flow cytometry can rapidly determine the number of cells in blood. For example, US patents US 6,114,173, US 4,882,284, and US 5,891,731 disclose methods for staining blood cells with dyes under non-hemolytic conditions to better distinguish platelets. Chinese patent application CN 101173921 discloses a specific staining agent for distinguishing platelets.

[0007] While optical methods can obtain accurate PLT measurement results, they require an additional dedicated detection channel in the blood cell analyzer, leading to longer measurement times per sample and reduced testing speed. Furthermore, this dedicated detection channel requires specialized diluents and dyes, resulting in higher equipment and testing costs, which hinders its clinical application.

[0008] Therefore, there is still a need to further improve platelet measurement methods. Summary of the Invention

[0009] In view of the above, the purpose of this invention is to provide a method for detecting platelets in blood samples without the need for a separate dedicated detection channel. This method utilizes a hemolytic channel, especially a conventional white blood cell detection channel, and a light source of a specific wavelength to accurately identify platelets in blood samples from blood shadow particles that are usually considered interference signals based on optical information.

[0010] A further objective of this invention is to provide an alarm or to identify and count reticulocytes in samples with abnormal reticulocyte content based on optical information, particularly fluorescence information.

[0011] Another object of the present invention is to compare and correct the platelet count detected by the present invention with the platelet count results detected by conventional impedance method.

[0012] Furthermore, an object of the present invention is to provide a blood testing system for implementing the above-described method.

[0013] To achieve the above objectives, the first aspect of the present invention provides a blood testing method, the method comprising:

[0014] A blood sample is treated with a first reagent to obtain a test sample. The first reagent includes a hemolysin, which lyses the red blood cells in the blood sample into fragments and keeps the morphology of the white blood cells and platelets in the blood sample substantially intact.

[0015] The particles in the test sample are passed one by one through the detection area of ​​the optical detection device, and the particles in the test sample are illuminated by the light source of the optical detection device to obtain the optical information of the test sample. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light.

[0016] The optical information of platelets in the test sample is obtained based on at least two types of light intensity information from the optical information of the test sample.

[0017] A second aspect of the present invention provides a blood testing method, the method comprising:

[0018] Blood samples are treated with a first reagent and a second reagent to obtain a test sample. The first reagent includes a hemolysin that breaks down red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample essentially intact. The second reagent includes a fluorescent dye.

[0019] The particles in the test sample are passed one by one through the detection area of ​​the optical detection device, and the particles in the test sample are illuminated by the light source of the optical detection device to obtain the optical information of the test sample. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light.

[0020] The optical information of reticulocytes in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample.

[0021] A third aspect of the present invention provides a blood testing method, the method comprising:

[0022] Prepare the first test sample containing the blood sample to be tested and the diluent;

[0023] The first test sample solution is flowed in a flow chamber with an electrode-equipped hole, and the electrical signal generated when particles in the first test sample solution pass through the hole is detected.

[0024] The first detection result of platelets in the first test sample is obtained based on the electrical signal.

[0025] When the first test result indicates that the platelets in the blood sample to be tested are abnormal...

[0026] Prepare a second test sample solution containing the blood sample to be tested and the diluent, or prepare a second test sample solution from the first test sample solution;

[0027] The second test sample solution is treated with a first reagent, the first reagent including a hemolytic agent, which at least completely lyses the mature red blood cells in the second test sample solution into fragments and keeps the cell morphology of the white blood cells and platelets in the second test sample solution substantially intact;

[0028] The particles in the second sample solution are passed one by one through the detection area of ​​the optical detection device, and the particles in the second sample solution are illuminated by the light source of the optical detection device to obtain the optical information of the second sample solution. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light.

[0029] The second detection result of platelets in the second test sample solution is obtained based on at least two light intensity information from the optical information of the second test sample solution.

[0030] A fourth aspect of the present invention provides a blood analysis system, the blood analysis system comprising:

[0031] A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle;

[0032] A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device, and the reagent supply unit provides a first reagent to the reaction chamber. The blood sample drawn by the sampling device is mixed with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that breaks down red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample substantially intact.

[0033] An optical detection apparatus includes a light source, a flow chamber, and at least two detectors. Particles of the test sample can flow within the flow chamber. Light emitted by the light source illuminates the particles in the flow chamber to generate optical information. The detectors collect the optical information. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light.

[0034] A data processing device electrically connected to the optical detection device and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: obtaining optical information of platelets in the test sample based on at least two types of light intensity information in the optical information of the test sample.

[0035] A fifth aspect of the present invention provides a blood analysis system, the blood analysis system comprising:

[0036] A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle;

[0037] A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device. The reagent supply unit provides a first reagent and a second reagent to the reaction chamber, thereby mixing the blood sample drawn by the sampling device with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample substantially intact. The second reagent includes a fluorescent dye.

[0038] An optical detection device includes a light source, a flow chamber, a scattered light detector, and a fluorescence detector. Particles of the test sample can flow within the flow chamber. Light emitted by the light source irradiates the particles in the flow chamber to generate optical information. The scattered light detector collects scattered light intensity information from the optical information, and the fluorescence detector collects fluorescence intensity information from the optical information. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light.

[0039] A data processing device electrically connected to the optical detection device and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: obtaining optical information of reticulocytes in the test sample based on fluorescence intensity information and scattered light intensity information in the optical information of the test sample.

[0040] This invention provides a novel method for detecting platelets, which can obtain accurate platelet information without adding additional detection channels or using additional specific detection reagents. The method uses a light source emitting light with a wavelength below 488nm and utilizes a hemolysis detection channel to detect hemolyzed blood samples, achieving complete differentiation of platelets from blood shadow particles through optical detection. Furthermore, this method can simultaneously obtain white blood cell analysis results and, with the use of fluorescent dyes, further obtain reticulocyte detection information. This allows for the simultaneous acquisition of white blood cell and even reticulocyte detection information in the same detection channel, thereby simplifying blood testing and reducing testing costs. Attached Figure Description

[0041] Figure 1 This is a schematic flowchart of a blood testing method according to the first aspect of the present invention;

[0042] Figure 2 This is a two-dimensional scatter plot of forward-scattered and side-scattered light obtained by treating blood samples with conventional hemolysing agents.

[0043] Figure 3 This is a schematic flowchart of a blood testing method according to a first embodiment of the first aspect of the present invention;

[0044] Figure 4 A two-dimensional scatter plot of forward-scattered light and side-scattered light obtained according to a first embodiment of the first aspect of the present invention;

[0045] Figure 5 A schematic flowchart of a blood testing method according to an example of a second embodiment of the first aspect of the present invention;

[0046] Figure 6A and Figure 6B Two-dimensional scatter plots obtained respectively, according to an example of a second embodiment of the first aspect of the present invention;

[0047] Figure 7 This is a schematic flowchart of a blood testing method according to another example of a second embodiment of the first aspect of the present invention;

[0048] Figure 8A and Figure 8B Two-dimensional scatter plots obtained respectively, according to another example of the second embodiment of the first aspect of the present invention;

[0049] Figure 9 A correlation comparison chart of platelet count PLT-1 measured by the method of the present invention and platelet count PLT-0 measured using a separate RET channel;

[0050] Figure 10 A comparison of the linear relationship between the reticulocyte relative count value RET-1 measured by the method of the present invention and the reticulocyte count value RET-0 measured using a separate RET channel;

[0051] Figure 11 Schematic diagram of the composition structure of the optical detection device provided in the embodiments of the present invention Figure 1 ;

[0052] Figure 12 Schematic diagram of the composition structure of the optical detection device provided in the embodiments of the present invention Figure 2 ;

[0053] Figure 13 This is a schematic flowchart of a blood testing method according to the second aspect of the present invention;

[0054] Figure 14 This is a schematic flowchart of a blood testing method according to a third aspect of the present invention;

[0055] Figure 15 This is a schematic diagram of the composition and structure of a blood cell analysis system provided in an embodiment of the present invention;

[0056] Figure 16 A schematic diagram of the optical detection device in the blood cell analysis system provided in an embodiment of the present invention;

[0057] Figure 17 This is a schematic diagram of an impedance detection device in a blood cell analysis system provided in an embodiment of the present invention. Detailed Implementation

[0058] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0059] Throughout this specification, unless otherwise specified, the terminology used herein should be understood as having the meaning commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In the event of any conflict, this specification shall prevail.

[0060] It should be noted that, in the embodiments of the present invention, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a method or apparatus that includes a list of elements includes not only the elements expressly stated, but also other elements not expressly listed, or elements inherent to implementing the method or apparatus. Without further limitations, an element defined by the phrase "comprising a..." does not exclude the presence of other related elements (e.g., steps in the method or units in the apparatus, where a unit may be a portion of circuitry, a portion of a processor, a portion of a program or software, etc.) in the method or apparatus that includes that element.

[0061] It should be noted that the terms "first," "second," and "third" used in the embodiments of this invention are merely to distinguish similar objects and do not represent a specific ordering of objects. It is understood that "first," "second," and "third" can be interchanged in a specific order or sequence where permissible. It should be understood that the objects distinguished by "first," "second," and "third" can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in an order other than those illustrated or described herein.

[0062] As mentioned earlier, the conventional method for detecting platelets is the electrical impedance method. However, this method is not accurate enough for certain blood samples. Therefore, optical detection methods combined with specific reagents have been reported for platelet detection in a separate detection channel. However, these methods increase the cost of the testing equipment and the overall testing expense.

[0063] US Patent 7,344,890 B2 discloses a method of treating blood samples containing interfering substances with a ghosting reagent to alter the scattering properties of red blood cells. This allows for the clear differentiation of platelets from red blood cells in a two-dimensional scatter plot by measuring the intensity and time-of-flight of the forward-scattered light from the cells in the sample. This method significantly alters the refractive index of red blood cells by causing them to lose heme. However, this method cannot effectively distinguish between large platelets and white blood cells. Furthermore, this method requires measuring the time-of-flight and cannot detect platelets solely based on optical information.

[0064] This invention proposes a method for detecting platelets using optical methods after hemolysis of blood samples. This method can completely distinguish platelets from lysed red blood cells obtained through hemolysis using optical information, and can also simultaneously obtain optical information of white blood cells. Furthermore, by using a fluorescent dye, optical information of reticulocytes can be further obtained to indicate the presence of reticulocytes in the blood sample.

[0065] The blood testing method and corresponding blood analysis system provided by the present invention will be described in detail below by way of example.

[0066] According to the blood detection method of the first aspect of the present invention, a light source emitting violet, blue, green, or yellow light, or a light source emitting light with a wavelength of less than 600 nanometers, especially less than 488 nanometers, is used for detection in order to completely distinguish platelets in the blood sample from other particles in the shadow region, so as to obtain an accurate platelet count.

[0067] Specifically, see Figure 1 The diagram illustrates a flowchart of a blood testing method according to a first aspect of the present invention. The blood testing method includes the following steps.

[0068] In step S11, the blood sample is treated with a first reagent to obtain a test sample. In this step, the first reagent includes a hemolysin to lyse red blood cells in the blood sample into fragments, while platelets retain their cellular morphology, and preferably white blood cells also retain their cellular morphology.

[0069] In step S12, optical information of each particle in the test sample is obtained through an optical detection device. Specifically, the particles in the test sample are passed one by one through the detection area of ​​the optical detection device, and the particles in the test sample are illuminated by the light source of the optical detection device to obtain the optical information of the test sample.

[0070] In step S13, the optical information of platelets in the test sample is obtained based on at least two types of light intensity information in the optical information of the test sample.

[0071] In this embodiment of the invention, the wavelength range of violet light is approximately 370nm to 435nm, the wavelength range of blue light is approximately 436nm to 500nm, the wavelength range of green light is approximately 501nm to 560nm, and the wavelength range of yellow light is approximately 561nm to 599nm. In one embodiment, the light source of the detection system of the present invention allows the emission of light with a wavelength in the range of approximately 370nm to 435nm, preferably in the range of 375nm to 420nm, and more preferably in the range of 400nm to 410nm. In another embodiment, the light source of the detection system of the present invention allows the emission of light with a wavelength in the range of approximately 436nm to 500nm, preferably in the range of 440nm to 500nm, and more preferably in the range of 445nm to 490nm. In yet another embodiment, the light source of the detection system of the present invention allows the emission of light with a wavelength in the range of approximately 501nm to 560nm, preferably in the range of 510nm to 550nm, and more preferably in the range of 510nm to 530nm.

[0072] In one embodiment, the light source of the detection system of the present invention can be configured to emit light with a wavelength of approximately 375 nm, 405 nm, 450 nm, or 520 nm.

[0073] Furthermore, the method according to the first aspect of the invention is implemented in a conventional leukocyte classification and / or counting channel without affecting the classification and / or counting of leukocytes. In the obtained scatter plot (see further examples detailed below), the leukocyte region is significantly separated from the platelet region. Therefore, the method of the present invention does not produce leukocyte interference with large platelets and platelet aggregates. Thus, the method of the present invention can simultaneously obtain at least three differential (e.g., monocytes, lymphocytes, and neutrophils), and even four differential (e.g., lymphocytes, monocytes, neutrophils, and eosinophils) leukocyte detection results when using fluorescent dyes, and can alarm or count reticulocytes.

[0074] Furthermore, the light source used in the blood detection method according to the first aspect of the present invention can be configured to emit light with a wavelength in the range of about 375 nm to 480 nm, especially light in the range of 405 nm to 480 nm, and more preferably light in the range of 440 nm to 480 nm.

[0075] According to a first embodiment of the first aspect of the present invention, the first reagent may contain a strong hemolytic agent capable of causing deep lysis of red blood cells, thereby completely lysing the red blood cells in the blood sample into fragments with light scattering characteristics significantly different from platelets. The strong hemolytic agent is not particularly limited. For example, such a strong hemolytic agent may be an alkyl glycoside, a triterpenoid saponin, a steroidal saponin, etc.

[0076] A specific strong hemolytic agent may be a glycoside compound having the general formula I:

[0077] R-(CH2) n -CH3 (I)

[0078] R is selected from the group consisting of monosaccharides, deoxymonosaccharides, and polysaccharides, and n is an integer from 5 to 17.

[0079] The aforementioned glycoside compounds can rapidly lyse red blood cells. Glycoside compounds are formed by the dehydration of the hemiacetal hydroxyl group of a sugar (or polysaccharide) and the hydroxyl group of an alkanol. The glycoside compound in the hemolytic agent of this invention can be a single compound or a mixture of two or more glycoside compounds conforming to the above general formula.

[0080] The concentration of the glycoside compound of general formula I in the hemolytic agent of the present invention varies depending on the properties of the selected glycoside, the reaction time, the reaction temperature, and the amount of other components used. The typical amount used is in the range of 0.025 g / L to 10 g / L, preferably 0.1 g / L to 5.0 g / L.

[0081] Furthermore, the first reagent may include a nonionic surfactant having general formula II:

[0082] R1-R2-(CH2CH2O) m -H (II)

[0083] Wherein, R1 is a C8-C23 alkyl group, and R2 is an -O-, Or -COO-, where m is an integer from 10 to 50; and

[0084] Optionally, at least one organic acid or a salt thereof, wherein the organic acid or salt thereof is selected from the group consisting of organic acids having at least one carboxyl or sulfonic acid group and their alkali metal salts.

[0085] Nonionic surfactants of formula II can bind to cell membranes to a certain extent, thus protecting the cell membranes of leukocytes and platelets from the effects of the aforementioned glycoside compounds and maintaining or essentially maintaining their cell morphology.

[0086] In this invention, compounds of general formula I and general formula II are used in combination to achieve rapid and deep lysis of red blood cells, and to protect platelet cell membranes in order to effectively detect platelets.

[0087] The ratio of compounds of general formula I and general formula II varies depending on the compounds selected. However, generally speaking, the ratio of compounds of general formula I to general formula II is 1:100 to 1:3, preferably 1:25 to 1:5, and more preferably 1:10 to 1:5.

[0088] According to a preferred embodiment of the present invention, the first reagent may further include at least one organic acid or its salt to improve the distinguishability of leukocyte side-scattered light.

[0089] The first reagent of the present invention may further include conventional additives. These additives may be selectively added as needed, such as (but not limited to) buffers, metal chelating agents, osmotic pressure regulators, preservatives, etc. These reagents are all commonly used in the art, as long as they do not interfere with the function of the above-mentioned components in the hemolytic agent of the present invention.

[0090] Specific embodiments of the first reagent of the present invention are described in the applicant’s prior international applications PCT / CN2019 / 084660 and PCT / CN2019 / 084648, the full contents of which are incorporated herein by reference.

[0091] The mixing ratio of the first reagent to the blood sample according to the first embodiment is not particularly limited. For example, the volume mixing ratio of the blood sample to the first reagent can be 1:40 to 1:60. The hemolysis reaction is carried out at a temperature such as 40 to 60°C for 15 to 100 seconds, preferably 40 to 80 seconds. The reaction temperature and time can be adjusted according to specific conditions.

[0092] Generally speaking, higher reaction temperatures and longer reaction times help to achieve a deeper degree of lysis of red blood cells.

[0093] In this invention, deep erythrocyte lysis (deep hemolysis) refers to the complete lysis of erythrocytes into fragments with light scattering properties significantly different from platelets, while platelets largely retain their cell morphology, and preferably leukocytes also largely retain their cell morphology. In a scatter plot of scattered light signals, platelets and deeply lysed erythrocyte fragments form two completely distinct particle groups. In contrast, conventional erythrocyte lysis in this invention refers to the use of conventional hemolysing agents. After the hemolysing agent reacts with the blood sample, in a scatter plot of scattered light signals, the lysed erythrocyte fragments and platelet particle groups may overlap, such as... Figure 2 As shown. Figure 2 Only the blood shadow area is shown, by Figure 2 It is known that the reticulocyte fragment region and the platelet region overlap to a certain extent, making it difficult to obtain an accurate platelet count.

[0094] In this invention, strong hemolytic agents, in particular, can completely lyse any reticulocytes that may be present into smaller fragments with light scattering characteristics significantly different from platelets. While conventional hemolytic agents, when used to treat blood cells, generally maintain the intact morphology of leukocytes and platelets and completely lyse mature erythrocytes into fragments, reticulocytes, due to their immaturity, are partially broken down. Some reticulocyte fragments may be similar in size to platelets, thus potentially causing overlap between reticulocyte fragments and platelet particle groups on a scatter plot of scattered light signals.

[0095] Therefore, in the first embodiment of the first aspect of the present invention, when red blood cells are deeply lysed using a strong hemolytic agent, the red blood cell fragment region and the platelet region can be clearly distinguished in a scatter plot composed of at least two types of scattered light intensity information obtained by optical detection, thereby achieving accurate and precise identification and counting of platelets. Furthermore, a white blood cell subset including at least monocytes, lymphocytes, and neutrophils can also be obtained.

[0096] Further reference Figure 3The diagram illustrates a flowchart of a specific blood testing method according to the first embodiment. In this specific blood testing method, firstly, in step S111, a blood sample is treated with a first reagent containing a strong hemolytic agent as described above to obtain a test sample. Next, in step S112, particles in the test sample are passed one by one through the flow chamber of an optical detection device to obtain optical information. In step S113, platelets are distinguished from other particles, especially completely lysed red blood cell fragments, based on forward and at least one other angle of scattered light intensity information. Furthermore, in step S114, optical information of white blood cells in the test sample is obtained based on the forward and side-scattered light intensity information in the optical information of the test sample, so as to distinguish white blood cell subpopulations based on the obtained white blood cell optical information to obtain a subpopulation of white blood cells including at least monocytes, lymphocytes, and neutrophils.

[0097] In one embodiment of the first aspect of the present invention, a blood sample is processed with decanyl glucoside to obtain a test sample, which is then measured using a blood cell analyzer (Mindray BC6200). The optical detection device of the Mindray BC6200 is modified, and the excitation wavelength of the laser light source is set to 405 nm. Forward and side-scattered light intensity information of particles in the test sample is acquired to plot a two-dimensional scatter plot of the test sample, such as... Figure 4 As shown. Figure 4 Only the blood shadow area is shown, from Figure 4 It can be seen that the red blood cell fragment region and the platelet region are highly distinguishable, and platelets can be clearly separated from the blood shadow region for effective statistical analysis.

[0098] In step S113, the scattered light intensity information at other angles includes at least one of side-scattered light intensity information, mid-angle scattered light intensity information, and high-angle scattered light intensity information. In one example, the at least two types of scattered light intensity information include forward-scattered light intensity information and side-scattered light intensity information.

[0099] The scattering angle of forward-scattered light signals can be approximately 1° to 10°. The scattering angle of mid-angle scattered light signals can be approximately 10° to 20°. The scattering angle of high-angle scattered light signals can be approximately 20° to 70°. The scattering angle of side-scattered light signals can be approximately 70° to 110°.

[0100] refer to Figure 5 The second embodiment of the present invention will be described below. Figure 5 A flowchart of a specific blood testing method in this second embodiment is shown.

[0101] In a second embodiment of the invention according to the first aspect, the hemolytic agent used in the first reagent is not particularly limited; it can be a strong hemolytic agent as described above, or a hemolytic agent that only routinely lyses red blood cells into fragments. In this second embodiment, the blood sample treated with the first reagent is further treated with a second reagent containing at least one fluorescent dye. See also Figure 5 In step S121, the blood sample is treated with a first reagent containing a hemolytic agent and a second reagent containing a fluorescent dye to obtain the test sample.

[0102] Specifically, in the second embodiment of the first aspect of the present invention, the first reagent may contain any hemolytic agent, as long as it can lyse red blood cells, and there is no particular limitation on the degree of hemolysis. It may be a strong hemolytic agent as described above, or a conventional hemolytic agent. Exemplary conventional hemolytic agents include quaternary ammonium cationic surfactants (such as tetradecyltrimethylammonium chloride), but the present invention is not limited thereto.

[0103] In the second embodiment of the first aspect of the present invention, since there are no special requirements for the degree of hemolysis, the time for the hemolysis reaction can be set to be shorter, for example, in the range of 15 to 30 seconds, especially around 20 seconds, so as to speed up the detection of the sample.

[0104] In step S122, optical information of the sample to be tested is also obtained through an optical detection device. Accordingly, in this step, in addition to the aforementioned scattered light intensity information, fluorescence intensity information is further obtained.

[0105] In step S123, the optical information of platelets in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information, especially the forward scattered light intensity information, in the optical information of the test sample, so as to distinguish platelets from other particles.

[0106] Furthermore, in step S124, the leukocyte subpopulations can be distinguished based on the side-scattered light intensity and fluorescence intensity information in the optical information of the test sample to obtain a leukocyte subpopulation including at least monocytes, lymphocytes and neutrophils and / or to identify immature granulocytes.

[0107] Furthermore, in step S125, the optical information of reticulocytes in the test sample can be obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample.

[0108] Furthermore, in step S126, when the optical information of the reticulocytes meets the preset conditions, a prompt indicating the presence of reticulocytes in the test sample can also be output.

[0109] In a second embodiment of the first aspect of the present invention, the second reagent may comprise a dye selected from membrane-specific dyes and mitochondrial-specific dyes, and / or a nucleic acid-specific dye.

[0110] The membrane-specific dye may be selected from one or more of DiA, DiD, DiI, DiO, DiR, DiS, FDA, Alexa Fluor 488, Super Fluor 488, and their derivatives. Preferably, the membrane-specific dye is Alexa Fluor 488.

[0111] The mitochondrial-specific dye may be selected from one or more of Janus Green B, MitoLite Red, Rhodamine 123, and the Mitotracker series, as well as their parent compounds. Preferably, the mitochondrial-specific dye is Mitotracker Deep Red or Mitotracker Red.

[0112] In this invention, the modified structures of the dye include commercially available modified structures and non-commercial modified structures. Based on the name, structure, etc., of the dye, those skilled in the art can identify modified structures (such as commercially available modified structures) based on known dyes from the prior art. Simultaneously, non-commercial modified structures can be obtained based on the parent structure and / or existing modified structures, and it is reasonably expected that these modified structures can achieve dyeing effects similar to their parent. All these modified structures fall within the protection scope of this invention.

[0113] In this invention, "membrane-specific dye" refers to a fluorescent dye capable of specifically staining the platelet membrane; similarly, "mitochondrial-specific dye" refers to a fluorescent dye capable of specifically staining the platelet mitochondria. When a blood sample is treated with a first reagent and a second reagent containing either a membrane-specific dye or a mitochondrial-specific dye, platelets and lysed red blood cells exhibit significantly different fluorescence characteristics. Therefore, by detecting fluorescence intensity and at least one selected from forward scattering intensity and side scattering intensity, especially by detecting fluorescence intensity and forward scattering intensity, platelets can be further distinguished from lysed red blood cell fragments, even when the blood sample is treated with conventional hemolytic agents.

[0114] In this invention, after reticulocytes in a blood sample are treated with a hemolytic agent, especially a strong hemolytic agent, the number of organelle particles released from the reticulocytes (RETs) is correlated with the number of RETs. Since reticulocytes are red blood cells containing nucleic acid, the addition of fluorescent dyes, especially nucleic acid-specific dyes, allows for specific staining of these particles.

[0115] Furthermore, our research revealed that in some blood samples containing a large number of reticulocytes, platelet detection using only two scattered light signals, particularly forward-scattered and side-scattered light signals, especially when using conventional hemolysins, can lead to interference from reticulocytes on platelets. A second reagent containing membrane- or mitochondrial-specific dyes can be used to trigger a reticulocyte alarm for this phenomenon. Specifically, particles associated with reticulocytes (i.e., organelle particles scattered after reticulocyte lysis) can be counted within a predetermined area of ​​a two-dimensional scatter plot of forward-scattered and fluorescence signals. When the count exceeds a predetermined value, a reticulocyte alarm can be triggered for further examination of the subject.

[0116] Therefore, in a specific example of the second embodiment of the first aspect of the present invention, it further includes: when the number of particles in a preset region of a scatter plot formed by forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample exceeds a predetermined threshold, it indicates the presence of reticulocytes in the test sample.

[0117] In one embodiment of the second embodiment of the first aspect of the present invention, a blood sample containing reticulocytes is processed with the mitochondrial dye Mitotracker Deep Red and a conventional hemolytic agent to obtain a test sample, which is then measured using a blood cell analyzer (Mindray BC6200). The optical detection device of the Mindray BC6200 is modified, and the excitation wavelength of the laser light source is set to 450 nm. The forward scattered light intensity, side scattered light intensity, and fluorescence intensity information of the particles in the test sample are acquired to plot a two-dimensional scatter plot of the test sample, as shown below. Figure 6A and Figure 6B . Figure 6A A two-dimensional scatter plot consisting of forward scattered light intensity information and fluorescence intensity information is shown, with only the blood shadow region shown; Figure 6B This shows a two-dimensional scatter plot composed of side-scattered light intensity information and fluorescence intensity information. Figure 6A It can be seen that the platelet's complete blood shadow region can be identified using forward scattering light intensity information and fluorescence intensity information. While the mature erythrocyte region and the reticulocyte region partially overlap, the presence of reticulocytes in the test sample can still be indicated by the number of particles in the preset region. Figure 6B It is known that this method has no effect on white blood cell count. White blood cell subsets can be distinguished by the intensity of side-scattered light and fluorescence intensity, and monocyte subset (MON), lymphocyte subset (LYM), neutrophil subset (NEU), and eosinophil subset (EOS) can be obtained.

[0118] In another specific example of the second embodiment of the first aspect of the present invention, the second reagent may comprise a nucleic acid-specific dye, particularly a dye specific to the nucleic acids of reticulocytes. This preferred embodiment allows staining of blood samples with nucleic acid dyes, thereby not only obtaining information about reticulocytes but also further distinguishing platelets from organelle particles scattered after reticulocyte lysis.

[0119] Therefore, this preferred embodiment of the invention includes the ability to further achieve effective determination of reticulocytes while detecting platelets.

[0120] refer to Figure 7 Describe the blood testing method used in this specific example. For example... Figure 7 As shown, steps S131, S132, and S133 of this method are the same as those described above. Figure 5 Steps S121 and S122 are the same, wherein the fluorescent dye of the second reagent includes a nucleic acid dye.

[0121] Furthermore, in step S133, the method can further distinguish reticulocyte-related particles, namely organelle particles after reticulocyte dissolution, by using forward scattering light intensity information and fluorescence intensity information. In other words, it can obtain the optical information of platelets and reticulocytes in the test sample.

[0122] Furthermore, after distinguishing between platelets and reticulocytes in step S133, particles associated with reticulocytes can be counted (step S134) to obtain the relative count value of reticulocytes.

[0123] In other words, according to the second embodiment of the first aspect of the present invention, platelets and reticulocytes in the test sample can be distinguished based on the scattered light intensity information, especially the forward scattered light intensity information and fluorescence intensity information, in the optical information of the test sample to obtain the optical information of platelets and reticulocytes in the test sample. Furthermore, the number of reticulocytes in the test sample can also be estimated based on the optical information of the reticulocytes.

[0124] In addition, these nucleic acid fluorescent dyes can also effectively stain leukocyte nuclei, and thus the fluorescence signal can be used to classify and detect leukocytes.

[0125] There are no particular limitations on the nucleic acid-specific dyes used in this invention. Commercially available nucleic acid fluorescent dyes and nucleic acid-specific fluorescent dyes disclosed in some patent applications can be used in this invention. Examples of commercially available nucleic acid fluorescent dyes include Thermofisher's SYTO series. Furthermore, fluorescent dyes disclosed in Chinese patent applications CN201010022414.6, CN200910109215.6 (anthocyanin-based dyes), and CN200810216864.1, etc., can all be used in this invention. The entire contents of the above patent documents are incorporated herein by reference.

[0126] The concentration range of the nucleic acid dye varies depending on the specific properties of the dye used and is not particularly limited, typically ranging from 0.002 ppm to 2000 ppm. A preferred concentration range is 0.03 ppm to 20 ppm.

[0127] The second reagent preferably further comprises an organic solvent. The organic solvent may be methanol, ethanol, glycerol, etc., but is not limited thereto.

[0128] In one specific embodiment, a blood sample containing reticulocytes was treated with the fluorescent dye SYTO9 (Thermofisher) and a conventional hemolysin to obtain a test sample. The sample was then analyzed using a blood cell analyzer (Mindray BC6200). The optical detection device of the Mindray BC6200 was modified, with the excitation wavelength of the laser light source set to 520 nm. The forward scattered light intensity, side scattered light intensity, and fluorescence intensity information of the particles in the test sample were acquired to plot a two-dimensional scatter plot of the test sample, as shown below. Figure 8A and Figure 8B . Figure 8A A two-dimensional scatter plot is shown, consisting of forward scattered light intensity information and fluorescence intensity information, with only the blood shadow region shown. Figure 8B This shows a two-dimensional scatter plot composed of side-scattered light intensity information and fluorescence intensity information. Figure 8A It can be seen that by using forward scattering light intensity information and fluorescence intensity information, the blood shadow region can be clearly divided into platelet region, mature red blood cell region, and reticulocyte region, thus enabling accurate platelet counting and relative reticulocyte counting. Figure 8B It is known that this method also has no effect on white blood cell count. White blood cell subsets can be distinguished by the intensity of side-scattered light and fluorescence intensity, and the monocyte subset (MON), lymphocyte subset (LYM), neutrophil subset (NEU), and eosinophil subset (EOS) can be obtained.

[0129] Furthermore, following the above method, a set of random blood samples were analyzed to obtain the platelet count (PLT-1) and reticulocyte relative count (RET-1) in the shadow region. Simultaneously, the corresponding platelet count (PLT-0) and reticulocyte count (RET-0) were measured using the RET independent channel of a blood cell analyzer. Correlation analysis was performed on the platelet count and reticulocyte count obtained from the two methods to obtain a fitted linear line, as shown below. Figure 9 and Figure 10 As shown, it can be seen that the platelet count PLT-1 and reticulocyte count PLT-1 measured by the method of the present invention have a good linear relationship with the platelet count PLT-0 and reticulocyte count PLT-0 measured by the PET single channel. Therefore, the separate reticulocyte channel can be eliminated, and the white blood cell channel can be used directly to achieve accurate platelet counting and alarm and relative counting of reticulocytes.

[0130] In a further preferred embodiment, the second reagent may contain one of a membrane- or mitochondrial-specific dye and a nucleic acid-specific dye to obtain a more accurate and precise platelet count, and simultaneously obtain a differential count of white blood cells and a relative count of reticulocytes.

[0131] In the aforementioned method using a second reagent containing a fluorescent dye, platelets can be distinguished from other particles in the sample based on information from fluorescence and forward scattering, allowing for platelet counting (and, in the case of nucleic acid dyes, relative reticulocyte counts). Furthermore, information from fluorescence and side scattering is used to obtain white blood cell classification and counts. Moreover, the intensity of fluorescence, forward scattering, and side scattering can be simultaneously used to obtain a three-dimensional scatter plot of volume distribution, thereby completing the classification and counting of each particle. Because the three-dimensional scatter plot reflects particle characteristics from multiple dimensions, it provides better differentiation between particle groups and yields more accurate results.

[0132] The blood detection method according to the first aspect of the present invention further includes counting platelets based on the obtained optical information of the platelets.

[0133] In a third embodiment of the first aspect of the present invention, the blood detection method of the first aspect of the present invention can further employ an optical detection device that eliminates the interference of laser pulse waves on platelet light signals after red blood cell lysis of blood samples according to the aforementioned first or second embodiment, so as to obtain a more accurate platelet count.

[0134] like Figure 11 and 12 As shown, Figure 11 and 12These are schematic diagrams illustrating the composition of a specific example of the optical detection device provided by the present invention. The optical detection device 200 includes an optical subsystem 1, a flow chamber 2, and a detector 3.

[0135] The optical subsystem 1 includes: a laser 11 (i.e., the light source of the present invention), a front optical assembly 12 including an optical isolator 121, and a rear optical assembly 13 including a beam deflector 131. The laser 11 is configured to emit a laser beam with a wavelength less than 600 nm, particularly less than 488 nm, or to emit violet, blue, green, or yellow light. The front optical assembly 12 is configured to perform pre-light processing on the laser beam, causing the pre-processed laser beam to converge at the beam deflector 131 in a second direction and then converge at the blood cell sample in the flow chamber 2 in a first direction, generating scattered light. The rear optical assembly 13 is disposed after the flow chamber 2 along the propagation direction of the laser beam and is configured to perform post-light processing on the scattered light and the laser beam converged at the beam deflector 131, so that the post-processed scattered light enters the first detector 3 for light intensity detection. The optical isolator 121 is configured to isolate the reflected light generated by the laser beam passing through the flow chamber and the rear optical assembly.

[0136] The first detector 3 of the optical detection device 200 is a forward-scattering light detector. The optical detection device 200 may also include a second detector 4. The second detector 4 may be a side-scattering light detector, or a mid-angle or high-angle scattered light detector. The optical detection device 200 may optionally also include a fluorescence detector 5.

[0137] In one embodiment, the front light assembly 12 further includes a collimating lens 122 disposed between the laser 11 and the optical isolator 121 along the propagation direction (optical axis direction) of the laser beam, configured to collimate the laser beam so that the laser beam becomes a parallel beam.

[0138] In one embodiment, the front light assembly 12 further includes a first light converging assembly 123 and a second light converging assembly 124. The first light converging assembly 123 is configured to perform a first focusing on the laser beam, so that the laser beam is focused in a first direction at the blood cell sample in the flow chamber and generates scattered light. The second light converging assembly 124 is configured to perform a second focusing on the laser beam, so that the laser beam is focused in a second direction at the directing aperture 131.

[0139] In one embodiment, the back light assembly 13 further includes a third converging assembly 132 and a pinhole aperture 133. The third converging assembly 132 is configured to perform a third focusing on the scattered light, so that the scattered light is converged at the pinhole aperture and enters the detector through the pinhole aperture for light intensity detection.

[0140] Specific embodiments of the optical inspection device are described in the applicant’s prior international applications PCT / CN2019 / 084660 and PCT / CN2019 / 084509, the full contents of which are incorporated herein by reference.

[0141] A second aspect of the present invention provides yet another method for blood testing. (See reference...) Figure 13 The flowchart of the method is shown, illustrating the steps of the method.

[0142] like Figure 13 As shown, in step S21, a blood sample is treated with a first reagent and a second reagent to obtain a test sample. The first reagent includes a hemolysin, which lyses the red blood cells in the blood sample into fragments and keeps the morphology of the white blood cells and platelets in the blood sample basically intact. The second reagent includes a fluorescent dye.

[0143] In step S22, particles in the test sample are passed one by one through the detection area of ​​the optical detection device, and the particles in the test sample are illuminated by the light source of the optical detection device to obtain optical information of the test sample. In this method, the light source is configured to emit light with a wavelength less than 600 nm, especially less than 488 nm, or to emit violet, blue, green, or yellow light.

[0144] Finally, in step S23, the optical information of reticulocytes in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample.

[0145] The second embodiment of the first aspect of the present invention described above can be applied to the blood detection method of the second aspect of the present invention, and will not be repeated here.

[0146] A third aspect of the present invention provides yet another method for blood testing. Reference is made below. Figure 14 This section further explains the blood testing method for the third aspect. Figure 14 A flowchart illustrating the method is shown.

[0147] In step S31, the blood sample to be tested is processed, for example, diluted with a diluent to prepare a first test sample.

[0148] In step S32, the electrical signal of the first test sample is obtained through an impedance detection device. Specifically, the first test sample is flowed in a flow chamber with an electrode-equipped orifice, and the electrical signal generated when particles in the first test sample liquid pass through the orifice is detected.

[0149] In step S33, the first detection result of platelets in the first test sample is obtained based on the electrical signal measured in step S32, that is, the impedance method detection result.

[0150] In step S34, it is determined whether there is an abnormality in the platelets indicated by the first test result. If the first test result indicates that there is no abnormality in the platelets in the blood sample to be tested, the first test result is reported. If the first test result indicates that there is an abnormality in the platelets in the blood sample to be tested, steps S35 to S39 are further performed.

[0151] In step S35, a second test sample is prepared by adding, for example, a diluent to the blood sample to be tested, or by preparing a second test sample from the first test sample. Further, in steps S36-S38, the second test sample is treated with a first reagent containing a hemolysin to obtain optical information of the second test sample, and a second detection result of the second test sample is obtained from at least two light intensity information in the optical information. That is, in steps S36-S38, the blood detection method provided by the first aspect of the present invention is used to obtain a second detection result of platelets in the second test sample, i.e., an optical detection result.

[0152] Furthermore, the abnormality is that the number of platelets in the blood sample is less than a predetermined threshold, that is, there are low platelet counts or platelet aggregation in the blood sample.

[0153] Furthermore, when the first detection result indicates that the platelets in the blood sample to be tested are abnormal, in step S39, the final detection result of the platelets in the blood sample to be tested is obtained based on the first detection result and the second detection result, or the second detection result is directly determined as the final detection result of the platelets in the blood sample to be tested.

[0154] This invention also provides another method for comparing and correcting the results of impedance platelet determination. In this method, regardless of whether there are platelet abnormalities in the blood sample, platelets in the blood sample are simultaneously detected in both the impedance detection channel and the hemolysis optical detection channel. The blood sample can be divided into two parts: one part is subjected to impedance detection using a conventional method, which will not be described in detail here; the other part is subjected to optical detection using the blood detection method described in the first aspect of this invention.

[0155] This method can simultaneously obtain platelet counting results from both impedance and optical methods.

[0156] Since the platelet detection method of this invention can obtain accurate platelet counts, under normal circumstances, the platelet counts measured by the two methods should be very close. When the difference between the platelet count result of the impedance method and the platelet count result of the optical method is greater than or equal to a predetermined threshold, it can be determined that the platelet content of the sample is abnormal, the result of the impedance method is inaccurate, and therefore the result of the optical method is selected to report the platelet count value, or the platelet count result of the optical method is used to correct the platelet count result of the impedance method; conversely, when the difference between the platelet count result of the impedance method and the platelet count result of the optical method is less than the predetermined threshold, it indicates that the detection result of the impedance method is accurate, and therefore the result of the impedance method and / or the result of the optical method can be reported.

[0157] A fourth aspect of the present invention provides a blood analysis system, the blood analysis system comprising:

[0158] A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle;

[0159] A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device, and the reagent supply unit provides a first reagent to the reaction chamber. The blood sample drawn by the sampling device is mixed with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that breaks down red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample substantially intact.

[0160] An optical detection apparatus includes a light source, a flow chamber, and at least two detectors. Particles of the test sample can flow within the flow chamber. Light emitted by the light source irradiates the particles in the flow chamber to generate optical information. The detectors collect the optical information. The light source is configured to emit light with a wavelength less than 600 nm, particularly less than 488 nm, or to emit violet, blue, green, or yellow light.

[0161] A data processing apparatus electrically connected to the optical detection apparatus and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing apparatus is configured to perform the steps of the method of the first or second aspect of the present invention as detailed above when the computer program is executed by the processor.

[0162] The processor mentioned above can be a central processing unit, or it can be other general-purpose processors, digital signal processors, application-specific integrated circuits, off-the-shelf programmable gate arrays or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. A general-purpose processor can be a microprocessor or any conventional processor.

[0163] The aforementioned computer-readable storage medium can be volatile memory or non-volatile memory, or both. Non-volatile memory can be read-only memory, programmable read-only memory, erasable programmable read-only memory, electrically erasable programmable read-only memory, magnetic random access memory, flash memory, magnetic surface memory, optical disk, or read-only optical disk; magnetic surface memory can be disk storage or magnetic tape storage. Volatile memory can be random access memory used as an external cache. By way of example, but not limitation, many forms of RAM are available, such as static random access memory, synchronous static random access memory, dynamic random access memory, synchronous dynamic random access memory, double data rate synchronous dynamic random access memory, enhanced synchronous dynamic random access memory, synchronous linked dynamic random access memory, and direct memory bus random access memory. The memory described in the embodiments of the present invention is intended to include these and any other suitable types of memory.

[0164] Figure 15 A specific blood analysis system according to the present invention is shown. The blood analysis system includes a first housing 100, a second housing 200, a sampling device 10, a sample preparation device 30, an optical detection device 50, a data processing device 70, and an output unit 90. In practical applications, the output unit 90 can be a user interface. In this embodiment, the optical detection device 50 and the data processing device 70 are disposed inside the second housing 200, respectively on both sides of the second housing 200. The sample preparation device 30 is disposed inside the first housing 100, and the output unit 90 and the sampling device 10 are disposed on the outer surface of the first housing 100.

[0165] The sampling device 10 has a sampling needle for collecting blood samples and delivering the collected blood samples to the sample preparation device 30. Depending on the implementation, the sampling device can collect multiple blood samples, provide them to different chambers of the sample preparation device for different processing, and subsequently perform different tests.

[0166] The sample preparation apparatus 30 has a reaction chamber and a reagent supply unit. The reagent supply unit stores reagents for reacting with blood samples (e.g., at least the aforementioned first reagent and an optional second reagent) and supplies the appropriate reagents to the reaction chamber as needed.

[0167] The sample preparation apparatus 30 may include at least one reaction chamber, wherein the at least one reaction chamber is configured to react the blood sample from the sampling unit with a reagent from the reagent supply unit to obtain a test solution containing a plurality of platelet particles, such that the platelet particles flow one by one through the flow chamber of the optical detection device.

[0168] The optical detection device 50 may include: the aforementioned optical subsystem with a light source, a flow chamber, and at least two detectors. The light source may emit wavelengths less than 600 nm, particularly less than 488 nm, or may emit violet, blue, green, or yellow light. The flow chamber allows blood particles, such as platelet particles, to pass through in a queue. The at least two detectors are used to collect optical information, particularly light intensity information, of the blood particles passing through the flow chamber.

[0169] In the apparatus of the present invention, the at least two detectors include a first detector, i.e., a forward-scattering light detector, for detecting the intensity of forward-scattered light from particles flowing in the flow chamber. This first optical detector is typically arranged on a straight line between the light source and the flow chamber, with the light source positioned on either side of the flow chamber. The at least two detectors also include a second detector. The second detector is arranged at an angle to the straight line between the light source and the flow chamber to detect the intensity of side-scattered light, mid-angle scattered light, or high-angle scattered light from particles flowing in the flow chamber.

[0170] The data processing device 70 is configured to detect blood (e.g., platelet) particles flowing through the flow chamber based on light intensity signals of at least two types of scattered light, and obtain the detection results of the corresponding blood particles.

[0171] The output unit 90 is configured to output the detection results corresponding to the blood (e.g., platelet) particles.

[0172] According to the aforementioned method of the present invention, when the first reagent contains a strong hemolytic agent, the at least two detectors include a forward-scattering light detector and a side-scattering light detector, and the data processing device is configured to further execute the various steps of the blood detection method of the first embodiment of the first aspect of the present invention when the computer program is executed by the processor.

[0173] In another embodiment, the reagent supply unit is configured to further provide a second reagent to the reaction cell, thereby mixing the blood sample drawn by the sampling device with the second reagent provided by the reagent supply unit in the reaction cell to prepare a test sample, wherein the second reagent includes a fluorescent dye, the optical detection device further includes a fluorescence detector, and the data processing device is configured to further execute the various steps of the blood detection method of the second embodiment of the first aspect of the present invention when the computer program is executed by the processor.

[0174] According to the method of the present invention, the data processing device of the blood analysis system utilizes fluorescence intensity information and forward scattered light intensity information to further distinguish platelets from reticulocytes or their fragments. Thus, the blood analysis system can accurately count platelets. The blood analysis system can also further obtain information on reticulocytes, and thereby issue an alarm for abnormalities in reticulocytes or count the reticulocytes.

[0175] The data processing device of this blood analysis system can also use fluorescence intensity, forward scatter light intensity and side scatter light intensity to obtain a three-dimensional scatter plot, thereby better distinguishing platelet particle groups from other particle groups and obtaining a more accurate platelet count.

[0176] See further Figure 16 The image shows a specific example of the optical detection device in the aforementioned blood analysis system. For example... Figure 16 As shown, the optical detection device has a light source 101, a beam shaping assembly 102, a flow chamber 103, and a forward-scattering light detector (i.e., a first detector) 104 arranged sequentially in a straight line. A dichroic mirror 106 is arranged at a 45° angle to the straight line on one side of the flow chamber 103. A portion of the side light emitted by the particles in the flow chamber 103 passes through the dichroic mirror 106 and is captured by a fluorescence detector (i.e., a third detector) 105, which is arranged at a 45° angle to the dichroic mirror 106 behind it; the other portion of the side light is reflected by the dichroic mirror 106 and captured by a side-scattering light detector (i.e., a second detector) 107, which is arranged at a 45° angle to the dichroic mirror 106 in front of it.

[0177] According to a fifth aspect of the present invention, a blood analysis system is provided, comprising:

[0178] A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle;

[0179] A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device. The reagent supply unit provides a first reagent and a second reagent to the reaction chamber, thereby mixing the blood sample drawn by the sampling device with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample substantially intact. The second reagent includes a fluorescent dye.

[0180] An optical detection device includes a light source, a flow chamber, a scattered light detector, and a fluorescence detector. Particles of the test sample can flow within the flow chamber. Light emitted by the light source irradiates the particles in the flow chamber to generate optical information. The scattered light detector collects the intensity of the scattered light from the optical information, and the fluorescence detector collects the fluorescence intensity information from the optical information. The light source is configured to emit light with a wavelength less than 600 nm, particularly less than 488 nm, or to emit blue, violet, green, or yellow light.

[0181] A data processing device electrically connected to the optical detection device and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the steps of the blood detection method of the second aspect of the present invention when the computer program is executed by the processor.

[0182] This invention provides a further blood analysis system. For example... Figure 17 As shown, in addition to the components described above, the blood analysis system may also include an impedance detection device 150. The impedance detection device 150 includes a flow chamber 151 with an aperture 152 having an electrode 153. The impedance detection device 150 detects the DC impedance generated when a particle in the test sample passes through the aperture 152 and outputs an electrical signal reflecting information about the particle's passage through the aperture.

[0183] Specifically, after aspirating a blood sample, the sampling device 10 is driven by its drive device and moves to the reaction chamber of the sample preparation device 30, injecting the aspirated blood sample into the reaction chamber. The delivery device 130 can also deliver the test sample, after being treated with diluent in the reaction chamber, to the impedance detection device 150, i.e., to the flow chamber 151. The impedance detection device 150 may also be provided with a sheath fluid chamber (not shown) to supply sheath fluid to the flow chamber 151. In the flow chamber 152, the test sample solution flows under the sheath fluid, and the orifice 152 causes the test sample solution to flow into a thin stream, allowing the particles (formed elements) contained in the test sample to pass through the orifice 152 one by one. The electrode 153 is electrically connected to a DC power supply 154, which provides DC current between the pair of electrodes 153. During the DC current supply by the DC power supply 154, the impedance between the pair of electrodes 153 can be detected. The resistance signal representing the impedance change is amplified by the amplifier 155 and then sent to the data processing device 70. The magnitude of the resistance signal corresponds to the volume (size) of the particle. Therefore, by processing the resistance signal through the data processing device 70, the platelet count result in the test sample can be obtained.

[0184] In this blood analysis system, the data processing device 70 is configured to implement the various steps of the blood detection method of the third aspect of the present invention, which will not be described in detail here.

[0185] The features mentioned above, as long as they are meaningful within the scope of this invention and do not contradict each other, can be combined arbitrarily. The advantages and features described for the blood analysis method of this invention are applied accordingly to the corresponding blood analysis system of this invention.

[0186] The above description is only a preferred embodiment of the present invention and does not limit the patent scope of the present invention. All equivalent structural transformations made under the inventive concept of the present invention using the contents of the present invention specification and drawings, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present invention.

Claims

1. A blood testing method, comprising: A blood sample is treated with a first reagent to obtain a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The particles in the test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the test sample are irradiated by the light source of the optical detection device to obtain the optical information of the test sample, wherein the light source is configured to emit light with a wavelength of less than 488 nm. The optical information of platelets in the test sample is obtained based on at least two types of light intensity information in the optical information of the test sample. and Platelet counts are performed based on the obtained optical information of the platelets. The hemolytic agent is a strong hemolytic agent, which completely lyses the red blood cells in the blood sample into fragments with light scattering properties significantly different from platelets. The at least two types of light intensity information include forward scattered light intensity information, and include at least one of side scattered light intensity information, mid-angle scattered light intensity information and high-angle scattered light intensity information, in order to distinguish platelets in the test sample from other particles and completely lysed red blood cell fragments.

2. A blood testing method, comprising: A blood sample is treated with a first reagent to obtain a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The particles in the test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the test sample are irradiated by the light source of the optical detection device to obtain the optical information of the test sample, wherein the light source is configured to emit violet light or blue light; The optical information of platelets in the test sample is obtained based on at least two types of light intensity information in the optical information of the test sample. and Platelet counts are performed based on the obtained optical information of the platelets. The hemolytic agent is a strong hemolytic agent, which completely lyses the red blood cells in the blood sample into fragments with light scattering properties significantly different from platelets. The at least two types of light intensity information include forward scattered light intensity information, and include at least one of side scattered light intensity information, mid-angle scattered light intensity information and high-angle scattered light intensity information, in order to distinguish platelets in the test sample from other particles, including completely lysed red blood cell fragments.

3. The blood detection method according to claim 1 or 2, wherein the light source is configured to emit light in the wavelength range of 375nm to 480nm.

4. The blood detection method according to claim 3, wherein the light source is configured to emit light in the wavelength range of 405nm to 480nm.

5. The blood detection method according to claim 3, wherein the light source is configured to emit light with a wavelength in the range of 440nm to 480nm.

6. The blood detection method according to claim 3, wherein the light source is configured to emit light with a wavelength of approximately 375 nm, 405 nm, or 450 nm.

7. The blood detection method according to claim 1, wherein the at least two types of light intensity information include forward-scattered light intensity information and side-scattered light intensity information, to distinguish platelets in the test sample from other particles, including completely lysed red blood cell fragments; and / or The method further includes: The optical information of leukocytes in the test sample is obtained based on the forward scattering light intensity information and the side scattering light intensity information in the optical information of the test sample, so as to distinguish leukocyte subpopulations based on the obtained leukocyte optical information to obtain leukocyte subpopulations including at least monocytes, lymphocytes and neutrophils.

8. A blood testing method, comprising: A blood sample is treated with a first reagent and a second reagent to obtain a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The second reagent includes a fluorescent dye. The particles in the test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the test sample are irradiated by the light source of the optical detection device to obtain the optical information of the test sample, wherein the light source is configured to emit light with a wavelength of less than 488 nm. The optical information of platelets in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample. and Platelet counts are performed based on the obtained optical information of the platelets.

9. A blood testing method, comprising: A blood sample is treated with a first reagent and a second reagent to obtain a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The second reagent includes a fluorescent dye. The particles in the test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the test sample are irradiated by the light source of the optical detection device to obtain the optical information of the test sample, wherein the light source is configured to emit violet light or blue light; The optical information of platelets in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample. and Platelet counts are performed based on the obtained optical information of the platelets.

10. The blood detection method according to claim 8 or 9, wherein the scattered light intensity information is forward scattered light intensity information.

11. The blood detection method according to claim 8 or 9, wherein the light source is configured to emit light in the wavelength range of 375nm to 480nm.

12. The blood detection method according to claim 11, wherein the light source is configured to emit light in the wavelength range of 405nm to 480nm.

13. The blood detection method according to claim 11, wherein the light source is configured to emit light in the wavelength range of 440nm to 480nm.

14. The blood detection method according to claim 11, wherein the light source is configured to emit light with a wavelength of approximately 375 nm, 405 nm, or 450 nm.

15. The blood testing method according to claim 8 or 9, wherein the method further comprises: Based on the lateral scattered light intensity and fluorescence intensity information in the optical information of the test sample, leukocyte subsets are distinguished to obtain leukocyte subsets including at least monocytes, lymphocytes and neutrophils and / or to identify immature granulocytes.

16. The blood testing method according to claim 8 or 9, wherein the method further comprises: The optical information of reticulocytes in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample; The fluorescent dyes mentioned therein include nucleic acid-specific dyes, which are nucleic acid-specific dyes for reticulocytes.

17. The blood testing method according to claim 16, wherein the method further comprises: When the optical information of the reticulocytes meets the preset conditions, a prompt is output indicating the presence of reticulocytes in the test sample.

18. The blood detection method according to claim 16, wherein the fluorescent dye comprises one selected from membrane-specific dyes and mitochondrial-specific dyes; The method further includes: When the number of particles in a preset region of a scatter plot formed by the forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample exceeds a predetermined threshold, it indicates the presence of reticulocytes in the test sample.

19. The blood testing method according to claim 17, The optical information of platelets and reticulocytes in the test sample is obtained based on the fluorescence intensity information and forward scattered light intensity information in the optical information of the test sample.

20. The blood testing method according to claim 19, wherein the method further comprises: The number of reticulocytes in the test sample is estimated based on the optical information of the reticulocytes.

21. A blood testing method, comprising: A blood sample is treated with a first reagent and a second reagent to obtain a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The second reagent includes a fluorescent dye. The particles in the test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the test sample are illuminated by the light source of the optical detection device to obtain the optical information of the test sample. The light source is configured to emit light with a wavelength of less than 488 nm, or to emit blue light or violet light. and The optical information of reticulocytes in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample; The fluorescent dyes mentioned therein include nucleic acid-specific dyes, which are nucleic acid-specific dyes for reticulocytes.

22. The blood testing method according to claim 21, wherein the method further comprises: Based on the lateral scattered light intensity and fluorescence intensity information in the optical information of the test sample, leukocyte subsets are distinguished to obtain leukocyte subsets including at least monocytes, lymphocytes and neutrophils and / or to identify immature granulocytes.

23. The blood testing method according to claim 21 or 22, wherein the method further comprises: When the optical information of the reticulocytes meets the preset conditions, a prompt is output indicating the presence of reticulocytes in the test sample.

24. The blood detection method according to claim 21 or 22, wherein the fluorescent dye comprises one selected from membrane-specific dyes and mitochondrial-specific dyes; The method further includes: When the number of particles in a preset region of a scatter plot formed by the forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample exceeds a predetermined threshold, it indicates the presence of reticulocytes in the test sample.

25. The blood testing method according to claim 21 or 22, Based on the fluorescence intensity information and forward scattering light intensity information in the optical information of the test sample, platelets and reticulocytes in the test sample are distinguished to obtain the optical information of reticulocytes in the test sample.

26. The blood testing method according to claim 25, wherein the method further comprises: The number of reticulocytes in the test sample is estimated based on the optical information of the reticulocytes.

27. A blood testing method, wherein the method comprises: Prepare the first test sample containing the blood sample to be tested and the diluent; The first test sample is flowed in a flow chamber with an electrode-equipped hole, and the electrical signal generated when particles in the first test sample pass through the hole is detected. The first detection result of platelets in the first test sample is obtained based on the electrical signal. When the first test result indicates that the platelets in the blood sample to be tested are abnormal... Prepare a second test sample containing the blood sample to be tested and a diluent, or prepare the second test sample from the first test sample; The second test sample is treated with a first reagent, the first reagent including a hemolytic agent, which lyses the red blood cells in the second test sample into fragments and keeps the cell morphology of the white blood cells and platelets in the second test sample intact; The particles in the second test sample that has undergone hemolysis treatment are passed one by one through the detection area of ​​the optical detection device and the particles in the second test sample are irradiated by the light source of the optical detection device to obtain the optical information of the second test sample. The light source is configured to emit light with a wavelength of less than 488 nm, or to emit blue light or violet light. and The second detection result of platelets in the second test sample is obtained based on at least two light intensity information from the optical information of the second test sample. The hemolytic agent is a strong hemolytic agent, which completely lyses the red blood cells in the blood sample into fragments with light scattering properties significantly different from platelets. The at least two types of light intensity information include forward scattered light intensity information, and include at least one of side scattered light intensity information, mid-angle scattered light intensity information and high-angle scattered light intensity information, in order to distinguish platelets in the test sample from other particles, including completely lysed red blood cell fragments.

28. A blood testing method, wherein the method comprises: Prepare the first test sample containing the blood sample to be tested and the diluent; The first test sample is flowed in a flow chamber with an electrode-equipped hole, and the electrical signal generated when particles in the first test sample pass through the hole is detected. The first detection result of platelets in the first test sample is obtained based on the electrical signal. When the first test result indicates that the platelets in the blood sample to be tested are abnormal... Prepare a second test sample containing the blood sample to be tested and a diluent, or prepare the second test sample from the first test sample; The second test sample is treated with a first reagent and a second reagent. The first reagent includes a hemolytic agent, which lyses the red blood cells in the second test sample into fragments and keeps the morphology of the white blood cells and platelets in the second test sample intact. The second reagent includes a fluorescent dye. The particles in the processed second test sample are passed one by one through the detection area of ​​the optical detection device and the particles in the second test sample are irradiated by the light source of the optical detection device to obtain the optical information of the second test sample. The light source is configured to emit light with a wavelength of less than 488 nm, or to emit blue light or violet light. and The second detection result of platelets in the second test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the second test sample.

29. The blood detection method according to claim 27 or 28, wherein the abnormality is that the number of platelets in the first test sample is less than a predetermined threshold.

30. The blood testing method according to claim 27 or 28, wherein the method further comprises: When the first detection result indicates that the platelets in the blood sample to be tested are abnormal, the final detection result of the platelets in the blood sample to be tested is obtained based on the first detection result and the second detection result, or the second detection result is determined as the final detection result of the platelets in the blood sample to be tested.

31. A blood analysis system, comprising: A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle; A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device, and the reagent supply unit provides a first reagent to the reaction chamber. The blood sample drawn by the sampling device is mixed with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. An optical detection apparatus includes a light source, a flow chamber, and at least two detectors. Particles of the test sample can flow within the flow chamber. Light emitted by the light source illuminates the particles in the flow chamber to generate optical information. The detectors collect the optical information. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light. A data processing device, electrically connected to the optical detection device and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to, when the computer program is executed by the processor, perform the following steps: obtaining optical information of platelets in the test sample based on at least two types of light intensity information in the optical information of the test sample; and counting platelets based on the obtained optical information of the platelets. The hemolytic agent is a strong hemolytic agent, which completely lyses the red blood cells in the blood sample into fragments with light scattering properties significantly different from platelets. The at least two types of light intensity information include forward scattered light intensity information, and include at least one of side scattered light intensity information, mid-angle scattered light intensity information and high-angle scattered light intensity information, in order to distinguish platelets in the test sample from other particles, including completely lysed red blood cell fragments.

32. The blood analysis system according to claim 31, wherein the light source is configured to emit light in the wavelength range of 375nm to 480nm.

33. The blood analysis system according to claim 32, wherein the light source is configured to emit light in the wavelength range of 405nm to 480nm.

34. The blood analysis system according to claim 32, wherein the light source is configured to emit light in the wavelength range of 440nm to 480nm.

35. The blood analysis system of claim 32, wherein the light source is configured to emit light with a wavelength of approximately 375 nm, 405 nm, or 450 nm.

36. The blood analysis system according to any one of claims 32 to 35, wherein the at least two detectors comprise a forward-scattering light detector and a side-scattering light detector, and the data processing device is configured to further perform the following steps when the computer program is executed by the processor: Based on the forward and side-scattered light intensity information in the optical information of the test sample, platelets in the test sample are distinguished from other particles, including completely lysed red blood cell fragments; and / or Based on the forward and side-scattered light intensity information in the optical information of the test sample, the white blood cells in the test sample are classified into at least monocytes, lymphocytes, and neutrophil subsets.

37. A blood analysis system, comprising: A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle; A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device. The reagent supply unit is configured to provide a first reagent and a second reagent to the reaction chamber, thereby mixing the blood sample drawn by the sampling device with the first and second reagents provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and maintains the morphology of white blood cells and platelets in the blood sample intact. The second reagent includes a fluorescent dye. An optical detection device includes a light source, a flow chamber, and at least two detectors. Particles of the test sample can flow within the flow chamber. Light emitted by the light source illuminates the particles in the flow chamber to generate optical information. The optical detection device collects this optical information. The light source is configured to emit light with a wavelength less than 488 nm, or to emit violet or blue light. The optical detection device includes a fluorescence detector. A data processing apparatus electrically connected to the optical detection apparatus and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing apparatus is configured to perform the following steps when the computer program is executed by the processor: The optical information of platelets in the test sample is obtained based on the fluorescence intensity information and scattered light intensity information in the optical information of the test sample; and platelets are counted based on the obtained optical information of platelets, and / or Based on the lateral scattered light intensity and fluorescence intensity information in the optical information of the test sample, leukocyte subsets are distinguished to obtain leukocyte subsets including at least monocytes, lymphocytes, and neutrophils, and / or immature granulocytes are identified, and / or The optical information of reticulocytes in the test sample is obtained from the fluorescence intensity information and scattered light intensity information in the optical information of the test sample. The fluorescent dyes mentioned therein include nucleic acid-specific dyes, which are nucleic acid-specific dyes for reticulocytes.

38. The blood analysis system according to claim 37, wherein the scattered light intensity information is forward scattered light intensity information.

39. The blood analysis system of claim 37, wherein the fluorescent dye comprises one selected from membrane-specific dyes and mitochondrial-specific dyes, and the data processing device is configured to further perform the following steps when the computer program is executed by the processor: When the number of particles in a preset region of a scatter plot formed by the forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample exceeds a predetermined threshold, it indicates the presence of reticulocytes in the test sample.

40. The blood analysis system according to any one of claims 37-39, wherein the data processing device is configured to further perform the following steps when the computer program is executed by the processor: Based on the forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample, the optical information of platelets and reticulocytes in the test sample is obtained.

41. The blood analysis system of claim 40, wherein the data processing device is configured to further perform the following steps when the computer program is executed by the processor: Based on the forward scattered light intensity and fluorescence intensity information in the optical information of the test sample, the optical information of platelets and reticulocytes in the test sample is obtained; and The number of reticulocytes in the test sample is estimated based on the optical information of the reticulocytes.

42. A blood analysis system, comprising: A sampling device having a suction tube with a suction nozzle and a driving device for driving the suction tube to quantitatively aspirate blood samples through the suction nozzle; A sample preparation apparatus includes a reaction chamber and a reagent supply unit. The reaction chamber receives a blood sample drawn by a sampling device. The reagent supply unit provides a first reagent and a second reagent to the reaction chamber, thereby mixing the blood sample drawn by the sampling device with the first reagent provided by the reagent supply unit in the reaction chamber to prepare a test sample. The first reagent includes a hemolysin that lyses red blood cells in the blood sample into fragments and keeps the morphology of white blood cells and platelets in the blood sample intact. The second reagent includes a fluorescent dye. An optical detection device includes a light source, a flow chamber, a scattered light detector, and a fluorescence detector. Particles of the test sample can flow within the flow chamber. Light emitted by the light source irradiates the particles in the flow chamber to generate optical information. The scattered light detector collects the scattered light intensity information from the optical information, and the fluorescence detector collects the fluorescence intensity information from the optical information. The light source is configured to emit light with a wavelength less than 488 nm, or to emit blue or violet light. A data processing device electrically connected to the optical detection device and including a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: obtaining optical information of reticulocytes in the test sample based on fluorescence intensity information and scattered light intensity information in the optical information of the test sample; The fluorescent dyes mentioned therein include nucleic acid-specific dyes, which are nucleic acid-specific dyes for reticulocytes.

43. The blood analysis system of claim 42, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: Based on the lateral scattered light intensity and fluorescence intensity information in the optical information of the test sample, leukocyte subsets are distinguished to obtain leukocyte subsets including at least monocytes, lymphocytes and neutrophils and / or to identify immature granulocytes.

44. The blood analysis system of claim 42 or 43, wherein the fluorescent dye comprises one selected from membrane-specific dyes and mitochondrial-specific dyes, and the data processing device is configured to perform the following steps when the computer program is executed by the processor: When the number of particles in a preset region of a scatter plot formed by the forward scattered light intensity information and fluorescence intensity information in the optical information of the test sample exceeds a predetermined threshold, it indicates the presence of reticulocytes in the test sample.

45. The blood analysis system according to claim 42 or 43, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: Based on the fluorescence intensity information and forward scattering light intensity information in the optical information of the test sample, platelets and reticulocytes in the test sample are distinguished to obtain the optical information of reticulocytes in the test sample.

46. ​​The blood analysis system of claim 45, wherein the data processing device is configured to perform the following steps when the computer program is executed by the processor: Based on the fluorescence intensity and forward scattered light intensity information in the optical information of the test sample, platelets and reticulocytes in the test sample are distinguished to obtain the optical information of the reticulocytes in the test sample; and The number of reticulocytes in the test sample is estimated based on the optical information of the reticulocytes.

Images