Method for classifying and counting white blood cells, hematology analyzer, and computer-readable storage medium

Abstract

The present invention provides a method for classifying and counting white blood cells, a hematology analyzer, and a computer-readable storage medium. The method includes: obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second lysing agent, wherein the ghost value in the second white blood cell histogram is less than the ghost value in the first white blood cell histogram, and the ghost value in the second white blood cell histogram is less than a threshold ghost value; determining the peak shape of the second white blood cell histogram; when the peak shape of the second white blood cell histogram is a double peak, classifying and counting white blood cells using the second white blood cell histogram; when the peak shape of the second white blood cell histogram is a single peak, classifying and counting white blood cells by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram.

Description

[0001] Specification Technical Field

[0002] The present invention generally relates to the technical field of blood analysis, and more specifically to a method for classifying and counting white blood cells, a hematology analyzer, and a computer-readable storage medium. Background Art

[0003] Currently, impedance methods are widely used for animal white blood cell counting and classification. Clinically, animal white blood cell counting and classification are of great significance for animal diagnosis and treatment. In the existing process of using impedance methods for animal white blood cell counting and classification, only counting and classification are performed after a single white blood cell reaction. This method still has some deficiencies. The fundamental reason lies in the particularity of animal blood cells, such as differences in morphology and volume, and different susceptibilities to environmental temperature, reagents, and reaction intensity. For example, under certain conditions, after reacting a human blood sample with a specific amount of hemolytic agent, the white blood cell volume histogram generated by collecting signals is less affected by blood shadows (red blood cell fragments, PLT, etc.), and the particle groups on the histogram are relatively spread out, which can achieve good white blood cell counting and classification effects.

[0004] However, for animal blood samples after reaction, it is difficult to balance counting and classification in the generated white blood cell histogram. If the amount of hemolytic agent is slightly less, the lymph particle part with a smaller volume in the generated animal white blood cell histogram is severely interfered by blood shadows, making it difficult to achieve accurate white blood cell counting and classification. At this time, the white blood cell particle clusters are usually relatively spread out. If the amount of hemolytic agent is slightly more, although the lymph particles with a smaller volume in the generated animal white blood cell histogram are less interfered by blood shadows and accurate white blood cell counting can be achieved, the white blood cells often have relatively dense particle cluster distributions, making it difficult to achieve accurate classification. On the other hand, the same amount of hemolytic agent may result in significantly different histograms for different animal white blood cell reactions. Therefore, in a single animal impedance white blood cell counting and classification, due to the particularity of animal blood cells and their sensitivity to external environmental conditions, there are certain challenges in animal white blood cell detection. Summary of the Invention

[0005] The present invention is proposed to solve at least one of the above problems. Specifically, on one hand, the present invention provides a method for classifying and counting white blood cells, the method comprising:

[0006] Obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a first hemolytic agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second hemolytic agent, wherein the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value;

[0007] Determine the peak type of the second white blood cell histogram;

[0008] When the peak type of the second white blood cell histogram is bimodal, perform white blood cell classification and counting using the second white blood cell histogram;

[0009] When the peak type of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification results and counting results of the second white blood cell histogram and the classification results of the first white blood cell histogram.

[0010] Another aspect of the present invention provides a method for white blood cell classification and counting, the method comprising:

[0011] Obtain a first white blood cell histogram of white blood cells in a blood sample treated with a hemolytic agent for a first treatment time and a second white blood cell histogram of white blood cells in a blood sample treated with the hemolytic agent for a second treatment time, wherein the second treatment time is greater than the first treatment time, the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value;

[0012] Determine the peak type of the second white blood cell histogram;

[0013] When the peak type of the second white blood cell histogram is bimodal, perform white blood cell classification and counting using the second white blood cell histogram;

[0014] When the peak type of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification results and counting results of the second white blood cell histogram and the classification results of the first white blood cell histogram.

[0015] Another aspect of the present invention provides a computer-readable storage medium, which includes a program that can be executed by a processor to implement the foregoing method.

[0016] Yet another aspect of the present invention provides a blood analyzer, comprising:

[0017] A reaction cell;

[0018] A sampling needle assembly for discharging a blood sample to be analyzed into the reaction cell;

[0019] A hemolytic agent pushing member for pushing a hemolytic agent into the reaction cell, the hemolytic agent including a first hemolytic agent and the second hemolytic agent;

[0020] A resistive detector for detecting white blood cells in a blood sample treated with the first hemolytic agent and generating a first signal, and detecting white blood cells in a blood sample treated with the second hemolytic agent and generating a second signal;

[0021] A processor for:

[0022] Obtaining a first white blood cell histogram based on the first signal and a second white blood cell histogram based on the second signal, wherein the ghost cell value in the second white blood cell histogram is less than the ghost cell value in the first white blood cell histogram, and the ghost cell value in the second white blood cell histogram is less than a threshold ghost cell value;

[0023] When the peak pattern of the second white blood cell histogram is bimodal, performing white blood cell classification and counting using the second white blood cell histogram;

[0024] When the peak pattern of the second white blood cell histogram is unimodal, performing white blood cell classification and counting by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram.

[0025] According to the method, blood analyzer and storage medium for white blood cell classification and counting according to the embodiments of the present invention, by obtaining the first white blood cell histogram and the second white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second lysing agent, and determining the manner of white blood cell counting and classification according to the peak pattern of the second white blood cell histogram. Since when the peak pattern of the second white blood cell histogram is bimodal and the ghost cell value in the second white blood cell histogram is less than the threshold ghost cell value, the ghost cell hardly interferes with the counting of the second white blood cell histogram. And because it is bimodal, the regional distribution of various white blood cell particle clusters in the second white blood cell histogram is relatively wide, so directly counting using the second white blood cell histogram can accurately obtain the classification and counting results of white blood cells in the blood. When the peak pattern of the second white blood cell histogram is unimodal, the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram are combined for white blood cell classification and counting. Since the distribution of white blood cell particle clusters is compact when the peak pattern of the second white blood cell histogram is unimodal, which is not conducive to classification, but its counting result is more accurate than that of the first white blood cell histogram because it is less affected by the ghost cell. And the first white blood cell histogram has a relatively wide distribution of white blood cell particle clusters due to being bimodal, which is conducive to classification. Therefore, by combining the classification result and counting result of the second white blood cell histogram determined to be unimodal and the classification result of the first white blood cell histogram for white blood cell classification and counting, more accurate white blood cell classification and counting results can be obtained. Therefore, the method according to the embodiments of the present invention can improve the accuracy of white blood cell counting and classification. Description of the Drawings

[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the accompanying drawings required for the description of the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can be obtained based on these drawings.

[0027] Figure 1 The schematic diagram of the first white blood cell histogram in an embodiment of the present invention is shown;

[0028] Figure 2 The schematic diagram of the second white blood cell histogram with a double-peak shape in an embodiment of the present invention is shown;

[0029] Figure 3 The schematic diagram of the second white blood cell histogram with a single-peak shape in an embodiment of the present invention is shown;

[0030] Figure 4 The flowchart of the method for white blood cell counting and classification in an embodiment of the present invention is shown;

[0031] Figure 5 The schematic diagram of the second white blood cell histogram with a double-peak shape in an embodiment of the present invention is shown;

[0032] Figure 6 The four-classification schematic diagram of the second white blood cell histogram with a double-peak shape in an embodiment of the present invention is shown;

[0033] Figure 7 The five-classification schematic diagram of the second white blood cell histogram with a double-peak shape in an embodiment of the present invention is shown;

[0034] Figure 8 The classification schematic diagram of the second white blood cell histogram with a single-peak shape in an embodiment of the present invention is shown;

[0035] Figure 9 The classification schematic diagram of the second white blood cell histogram with a single-peak shape in another embodiment of the present invention is shown;

[0036] Figure 10 The schematic diagram of delimiting the blood shadow area in the first white blood cell histogram in an embodiment of the present invention is shown;

[0037] Figure 11 The schematic diagram of delimiting the blood shadow area in the first white blood cell histogram in another embodiment of the present invention is shown;

[0038] Figure 12 The first white blood cell histogram in an embodiment of the present invention is shown Figure 3 The classification schematic diagram;

[0039] Figure 13 Shows a flowchart of a method for realizing four-category white blood cell classification in an embodiment of the present invention;

[0040] Figure 14 Shows a flowchart of a method for realizing five-category white blood cell classification in an embodiment of the present invention;

[0041] Figure 15 Shows a classification schematic diagram of a second white blood cell histogram with a single peak shape in another embodiment of the present invention;

[0042] Figure 16 Shows the first white blood cell histogram in another embodiment of the present invention Figure 3 Classification schematic diagram;

[0043] Figure 17 Shows a flowchart of a method for white blood cell counting and classification in still another embodiment of the present invention;

[0044] Figure 18 Shows a schematic block diagram of an exemplary electronic device for implementing a method and apparatus for white blood cell classification and counting according to an embodiment of the present invention;

[0045] Figure 19 Shows a schematic block diagram of a blood analyzer in an embodiment of the present invention. Detailed implementation manners

[0046] In order to make the objectives, technical solutions and advantages of the present invention more apparent, exemplary embodiments according to the present invention will be described in detail below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments of the present invention. It should be understood that the present invention is not limited by the exemplary embodiments described herein. Based on the embodiments of the present invention described herein, all other embodiments obtained by those skilled in the art without creative efforts shall fall within the protection scope of the present invention.

[0047] In the following description, numerous specific details are given to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known to those skilled in the art are not described.

[0048] It should be understood that the present invention can be implemented in different forms and should not be construed as limited to the embodiments presented herein. On the contrary, providing these embodiments will make the disclosure thorough and complete, and will fully convey the scope of the present invention to those skilled in the art.

[0049] The purpose of the terms used herein is only to describe specific embodiments and is not a limitation of the present invention. As used herein, the singular forms "a", "an" and "the" are also intended to include the plural forms unless the context clearly dictates otherwise. It should also be understood that the terms "comprising" and / or "including", when used in this specification, specify the presence of the stated features, integers, steps, operations, elements and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components and / or groups. As used herein, the term "and / or" includes any and all combinations of the associated listed items.

[0050] To thoroughly understand the present invention, detailed structures will be presented in the following description to illustrate the technical solutions proposed by the present invention. The alternative embodiments of the present invention are described in detail below. However, in addition to these detailed descriptions, the present invention may also have other embodiments.

[0051] In order to solve the technical problems in blood detection, in a single white blood cell counting and classification, due to the particularity of blood cells and their sensitivity to external environmental conditions, there are certain challenges in white blood cell detection and poor accuracy. Embodiments of the present invention provide a method for white blood cell classification and counting. The method includes: obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second lysing agent, wherein the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value; determining the peak type of the second white blood cell histogram; when the peak type of the second white blood cell histogram is bimodal, performing white blood cell classification and counting with the second white blood cell histogram; when the peak type of the second white blood cell histogram is unimodal, combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram to perform white blood cell classification and counting.

[0052] Through the above method, after obtaining two white blood cell counts under different lysing agents, generating a first white blood cell histogram and a second white blood cell histogram, and comprehensively analyzing the method of white blood cell counting and classification according to the peak type characteristics of the second white blood cell histogram, thereby improving the accuracy of white blood cell counting and classification.

[0053] Specifically, the method for white blood cell classification and counting of the present application will be described in detail below with reference to the accompanying drawings. Without conflict, the features in the following embodiments and implementation manners can be combined with each other.

[0054] In one embodiment, as Figure 4 shown, the method for white blood cell counting and classification in embodiments of the present invention includes the following steps S401 to S404:

[0055] First, in step S401, a first white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second lysing agent are obtained, where the ghost value in the second white blood cell histogram is less than the ghost value in the first white blood cell histogram, and the ghost value in the second white blood cell histogram is less than a threshold ghost value.

[0056] The above blood sample can be any animal or human blood sample. Among them, the animal blood sample can be the blood sample of a dog, a cat, etc. The blood sample includes white blood cells. In this embodiment, the method of the embodiment of the present invention is mainly explained and illustrated by taking the animal blood sample as an example, but it can be imagined that the method of the present invention can also be applied to human blood samples. In this article, both the first white blood cell histogram and the second white blood cell histogram are obtained based on impedance measurement of the blood sample.

[0057] The blood sample can be treated with a diluent, a lysing agent, etc. Among them, the diluent is an isotonic solution with acid-base buffering effect, appropriate ionic strength and conductivity. For example, the diluent mainly contains hypoxanthine or xanthine compounds or their salts, or it can also be other diluents that can play the above roles. The role of the lysing agent is to lyse red blood cells for white blood cell classification and counting. The lysing agent includes surfactants, which specifically can include cationic surfactants and non-ionic surfactants. For example, the lysing agent can mainly contain quaternary ammonium salt ionic surfactants, or it can also be any other surfactant that can play the above roles.

[0058] For the same blood sample, such as a human blood sample or an animal blood sample, especially an animal blood sample, under different reaction conditions, such as different amounts of lysing agent, diluent, temperature, and stirring intensity, etc., generally two types of white blood cell histograms can be generated. For example Figure 1 the first white blood cell histogram shown, and as Figure 2 and Figure 3 the second white blood cell histogram shown.

[0059] In this embodiment, other reaction conditions (such as the amount of diluent, temperature, and stirring intensity, etc.) are mainly kept constant, and different reaction conditions are obtained by changing at least one of the type, dose, concentration, etc. of the lysing agent, so as to generate different white blood cell histograms.

[0060] In one example, white blood cells in a blood sample treated with a first hemolytic agent are detected based on impedance method to obtain a first white blood cell histogram, and white blood cells in a blood sample treated with a second hemolytic agent are detected based on impedance method to obtain a second white blood cell histogram. Among them, the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value. This threshold blood shadow value can be reasonably set based on prior experience. For example, when the blood shadow value in the second white blood cell histogram is less than this threshold blood shadow value, the influence on the white blood cell count result of the second white blood cell histogram can be ignored, so as to ensure the accuracy of the white blood cell count result of the second white blood cell histogram. Optionally, a blood shadow area can be delimited in the second white blood cell histogram, the area of this blood shadow area can be calculated or the count result of the blood shadows included in this blood shadow area can be calculated as the blood shadow value, and then it is compared with the threshold blood shadow value to determine whether the blood shadow value in the obtained second white blood cell histogram meets the requirement of being less than this threshold blood shadow value.

[0061] The first hemolytic agent and the second hemolytic agent can be hemolytic agents of different types or the same hemolytic agent. In this embodiment, the case where the first hemolytic agent and the second hemolytic agent are the same hemolytic agent is mainly used as an example to explain and illustrate the method of the present invention, but this is not intended to limit the scope of the present invention.

[0062] Optionally, the dose of the second hemolytic agent is greater than the dose of the first hemolytic agent, so that the number of blood shadows in the blood sample treated with the first hemolytic agent is greater than the number of blood shadows in the blood sample treated with the second hemolytic agent, so as to facilitate obtaining the aforementioned second white blood cell histogram and first white blood cell histogram.

[0063] In one example, as Figure 13 and Figure 14 shown, the method for obtaining the first white blood cell histogram and the second white blood cell histogram includes: first adding the first hemolytic agent to the blood sample to obtain a test solution, and obtaining the first white blood cell histogram; then adding the second hemolytic agent to the test solution to obtain the second white blood cell histogram, where the first hemolytic agent and the second hemolytic agent can be the same hemolytic agent. For example, after each hemolytic agent treats the blood sample, white blood cells in the blood sample are detected based on impedance method to obtain the first white blood cell histogram and the second white blood cell histogram. Since the blood sample already includes the first hemolytic agent when adding the second hemolytic agent, the blood shadow value in the blood sample treated with the second hemolytic agent is less than that of the first hemolytic agent. Therefore, by detecting the blood sample treated with the second hemolytic agent based on impedance method, the second white blood cell histogram can be obtained.

[0064] Specifically, the dosages of the first hemolytic agent and the second hemolytic agent added can be reasonably set according to actual needs, or can also be reasonably set according to prior experience. For example, based on prior experience, such as after several experiments, for example, 100 experiments, after adding different dosages of the first hemolytic agent to a blood sample, a corresponding white blood cell histogram is generated each time. By analyzing the characteristics of the white blood cell histogram generated each time, it is found that the white blood cell histogram generated within the threshold dosage range of the first hemolytic agent generally meets the requirements of the first white blood cell histogram in the embodiments of the present invention (for example, the first white blood cell histogram in the embodiments of the present invention needs to meet the following requirements: the blood shadow value included in the first white blood cell histogram is greater than or equal to the threshold blood shadow value, and its peak shape is a double peak). Thus, the range interval of the dosage of the first hemolytic agent added can be generally determined, and then the dosage of the first hemolytic agent added can be determined.

[0065] Similarly, based on prior experience, such as after several experiments, for example, 100 experiments, the second hemolytic agent is added to the blood sample treated with a predetermined dosage (the predetermined dosage is within the threshold dosage range of the aforementioned first hemolytic agent). A corresponding white blood cell histogram is generated each time. By analyzing the characteristics of the white blood cell histogram generated each time, it is found that the white blood cell histogram generated within the threshold dosage range of the second hemolytic agent generally meets the requirements of the second white blood cell histogram in the embodiments of the present invention (for example, the second white blood cell histogram in the embodiments of the present invention needs to meet the following requirements: the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than the threshold blood shadow value). Thus, the range interval of the dosage of the second hemolytic agent added can be generally determined, and then the dosage of the second hemolytic agent added can be determined.

[0066] In another example, obtaining the first white blood cell histogram of white blood cells in the blood sample treated with the first hemolytic agent and the second white blood cell histogram of white blood cells in the blood sample treated with the second hemolytic agent includes: adding at least three times of hemolytic agents to the same blood sample in sequence, each time the hemolytic agent added includes at least one of the first hemolytic agent and the second hemolytic agent, and generating a corresponding white blood cell histogram after each addition of the first hemolytic agent and / or the second hemolytic agent; selecting two white blood cell histograms generated successively as the first white blood cell histogram and the second white blood cell histogram respectively. For example, selecting two white blood cell histograms that meet the following requirements successively as the first white blood cell histogram and the second white blood cell histogram. Among them, the blood shadow value included in the first white blood cell histogram is greater than or equal to the threshold blood shadow value, and its peak shape is a double peak. The blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than the threshold blood shadow value.

[0067] It is worth mentioning that the same blood sample can be sampled into the same reaction cell or counting cell, and then the above reactions of adding the first hemolytic agent and the second hemolytic agent are carried out. Alternatively, the same blood sample can be separately sampled and added into at least two reaction cells or counting cells, and then the above reactions of adding the first hemolytic agent and the second hemolytic agent are carried out. However, no matter which method is used, the above-mentioned first white blood cell histogram and second white blood cell histogram can be finally obtained.

[0068] It is worth mentioning that the characteristics of blood cells inside the blood samples of different animal types (such as dogs, cats, pigs, sheep, cows, etc.) may be different. It is very likely that the dosages of the first hemolytic agent and the second hemolytic agent to be added to the blood samples of different animal types will vary. Therefore, the dosages of the first hemolytic agent and the second hemolytic agent can also be reasonably selected according to the animal type of the blood sample.

[0069] In one example, when this method is applied to a blood analyzer of an animal model, the blood analyzer is also provided with a detection mode selection module, such as a detection mode button set on the blood analyzer. Before detecting the blood sample of the embodiment of the present invention, the method further includes: selecting a predetermined detection mode to detect the blood sample according to the animal type to which the blood sample belongs, so as to obtain the first white blood cell histogram and the second white blood cell histogram, where the predetermined detection mode may include a cat blood sample detection mode, a dog blood sample detection mode, or other types of animal blood sample detection modes.

[0070] In one example, taking the blood sample of a cat as an example, after adding the same hemolytic agent to the same blood sample at least twice in sequence, the obtained white blood cell histogram may include a white blood cell histogram with a blood shadow (the blood shadow value is greater than or equal to the threshold blood shadow value) but a single-peak shape, and also includes a white blood cell histogram with a blood shadow (the blood shadow value is greater than or equal to the threshold blood shadow value) but a double-peak shape. In this embodiment, in order to facilitate the classification of the first white blood cell histogram, the white blood cell histogram with a blood shadow (the blood shadow value is greater than or equal to the threshold blood shadow value) but a double-peak shape is selected as the first white blood cell histogram.

[0071] Therefore, as continued in Figure 4 shown, the method of the embodiment of the present invention further includes step S402 of determining the peak shape of the second white blood cell histogram.

[0072] The peak shape of the second white blood cell histogram can be determined by any suitable method. The peak shape refers to, for example, the number of peaks included in the second white blood cell histogram. For example, if the number of peaks is 1, its peak shape is a single peak, and if the number of peaks is 2, its peak shape is a double peak, and so on.

[0073] In a specific example, the method for determining the peak type of the second white blood cell histogram includes: determining the peak type of the second white blood cell histogram according to the number of peak points in the second white blood cell histogram, where if the number of peak points is 2 and the distance between the two peak points is greater than a preset distance, then the second white blood cell histogram is bimodal; otherwise, the second white blood cell histogram is unimodal. This preset distance can be reasonably set according to the characteristics of the actual white blood cell histogram. In this article, the preset distance can refer to the volume size between adjacent peak points. For example, the preset distance can be about 40 fL, or other suitable distances, such as Figure 5 In the second white blood cell histogram shown, it is determined that there are two peak points A and B, and the distance between peak point A and peak point B is approximately 50 fL, which is greater than the preset distance S. Therefore, it can be determined that Figure 5 the peak type of the second white blood cell histogram shown is bimodal.

[0074] It is worth mentioning that in this article, when determining the peak points of the white blood cell histogram (such as the first white blood cell histogram and the second white blood cell histogram), there are at least a predetermined number of consecutive point ordinate values on both the left and right sides of the peak point that are less than the ordinate value at the peak point. This predetermined number is reasonably set according to actual needs, such as at least 10 points, 20 points, 30 points, 40 points, or other suitable numbers of points, etc. Exemplarily, it can start from the minimum volume (V min ) or the maximum volume (V max ) of the white blood cell histogram and traverse each point on the white blood cell histogram curve. For example, starting from the minimum volume and traversing each point on the white blood cell histogram curve in the direction of increasing volume, when the ordinate value of the point gradually increases to the maximum value, and then as the volume continues to increase, the ordinate values of at least a predetermined number of consecutive points start to decrease from the maximum value, so that the maximum value can be determined as the peak point. Similarly, it can also start from the minimum volume and traverse each point on the white blood cell histogram curve in the direction of decreasing volume. When the ordinate value of the point gradually increases to the maximum value, and then as the volume continues to decrease, the ordinate values of at least a predetermined number of consecutive points start to decrease from the maximum value, so that the maximum value can be determined as the peak point.

[0075] Continue to refer to Figure 4 , in step S403, when the peak type of the second white blood cell histogram is bimodal, white blood cell classification and counting are performed using the second white blood cell histogram.

[0076] In a specific example, as Figure 13 shown, when the peak type of the second white blood cell histogram is bimodal, the second white blood cell histogram determined to be bimodal is classified into four categories, and the classification result of this four-category classification is used as the white blood cell four-category classification result of the final blood sample.

[0077] It is worth mentioning that in this article, the white blood cell histogram is classified into three categories, four categories, or five categories, etc. Among them, the three-category classification may refer to classifying white blood cells into the first type of white blood cells, the second type of white blood cells, and the third type of white blood cells. The four-category classification may refer to classifying white blood cells into the first type of white blood cells, the second type of white blood cells, the third type of white blood cells, and the fourth type of white blood cells. The five-category classification may refer to classifying white blood cells into the first type of white blood cells, the second type of white blood cells, the third type of white blood cells, the fourth type of white blood cells, and the fifth type of white blood cells. Among them, for different types of blood samples, the white blood cells may contain different types of white blood cells. For example, for animal white blood cells, starting from the smallest volume, they are the first to fifth types of white blood cells in sequence. Among them, the first type of white blood cells may be lymphocytes, the second type of white blood cells may be monocytes, the third type of white blood cells may be neutrophils, the fourth type of white blood cells may be eosinophils, and the fifth type of white blood cells may be basophils.

[0078] Any suitable method can be used for the four-category classification of the second white blood cell histogram. For example, as Figure 6 shown, the method for the four-category classification of the second white blood cell histogram may include the following steps A1 to A4:

[0079] First, in step A1, according to the trough point C between the two peak points A and B in the second white blood cell histogram, the first dividing line 1 between the first type of white blood cells and the second type of white blood cells is determined. Among them, the volume of the first type of white blood cells is smaller than the volume of the second type of white blood cells. The area in the second white blood cell histogram with a volume smaller than the volume of the first dividing line is the first type of white blood cells (such as lymphocytes (LYM) shown as Figure 6 shown).

[0080] The two peak points A and B in the second white blood cell histogram can be determined by the method mentioned above, and the trough point C can be determined by any suitable method. For example, the trough point C is the minimum value point corresponding to the minimum ordinate value between the peak point A and the peak point B.

[0081] Next, in step A2, continue as Figure 6As shown, a second boundary line 2 between the second type of white blood cells (e.g., monocytes (MON)) and the third type of white blood cells (e.g., neutrophils (NEU)) is determined according to the first boundary line 1, wherein the second boundary line and the first boundary line are spaced apart by a first predetermined volume, and the volume corresponding to the second boundary line is greater than the volume corresponding to the first boundary line;

[0082] This first predetermined volume Smon can be reasonably set according to prior experience. For example, under specific reaction conditions, reaction temperature, and reagent (including lysing agent and diluent) dosages, after multiple detections, the volume between the trough point and the actual second boundary line under these specific conditions, especially under a specific lysing agent dosage, is obtained to determine the first predetermined volume Smon. For example, Smon is about 20 fL. Among them, the positions of the trough points and the values of the first predetermined volume Smon will also be different under different reaction conditions, reaction temperatures, and reagent (including lysing agent and diluent) dosages, and can be reasonably adjusted according to the actual situation.

[0083] Next, in step A3, starting from the maximum volume Vmax of the second white blood cell histogram (e.g., Vmax = 250 fL), the second critical point D on the curve of the second white blood cell histogram where the slope is first greater than the second threshold slope K is searched in the direction of decreasing volume;

[0084] Among them, the maximum volume Vmax may refer to the end position of white blood cells in the white blood cell histogram. Usually, the curve of the initial white blood cell histogram on the right side of the first peak point A shows an upward trend. The slope of the points on the curve of the second white blood cell histogram within a predetermined segment starting from the maximum volume Vmax in the direction of decreasing volume is less than or equal to 0. Therefore, the value of the second threshold slope K is set to be less than zero. Specifically, the value of the second threshold slope K can be set according to the actual situation.

[0085] Next, in step A4, based on the second critical point, a third boundary line between the third type of white blood cells and the fourth type of white blood cells is determined. This third boundary line is a straight line passing through the second critical point D and perpendicular to the horizontal axis of the second white blood cell histogram, thereby realizing the four-classification of white blood cells. Among them, the area between the second boundary line and the third boundary line is the third type of white blood cells, such as neutrophils NEU, and the area between the third boundary line and Vmax is the fourth type of white blood cells (e.g., eosinophils EOS).

[0086] In another specific example, as Figure 14 shown, when the peak shape of the second white blood cell histogram is a double peak, the second white blood cell histogram determined to be a double peak is classified into five classes, and the classification result of this five-classification is used as the white blood cell five-classification result of the final blood sample.

[0087] Any suitable method can be adopted for the five-classification of the second white blood cell histogram. For example, as Figure 7 shown, the method for the five-classification of the second white blood cell histogram may include steps A1 to A4 of the aforementioned four-classification, and further include: determining a fourth boundary line between the fourth type of white blood cells and the fifth type of white blood cells according to the third boundary line, wherein the fourth boundary line is spaced from the third boundary line by a second predetermined volume S baso , and the volume corresponding to the fourth boundary line is greater than the volume corresponding to the third boundary line. Then, the fourth type of white blood cells is the region on the second white blood cell histogram located between the third boundary line and the fourth boundary line, and the fifth type of white blood cells (such as basophils (BASO)) is the region on the second white blood cell histogram with a volume greater than the volume corresponding to the fourth boundary line, that is, the region between the fourth boundary line and the maximum volume.

[0088] The second predetermined volume S baso can be reasonably set according to prior experience. For example, the second predetermined volume S baso is about 55 fL, or other suitable volumes. Among them, due to different reaction conditions, the value of this predetermined volume may also change. Therefore, the above numerical values are only examples and are not intended to limit the scope of the present invention.

[0089] It is worth mentioning that in this article, the rectangular coordinate system where the white blood cell histogram is located has the cell volume size as the abscissa and the number of cell particles with different volumes as the ordinate. Among them, the first boundary line, the second boundary line, the third boundary line, and the fourth boundary line are all perpendicular to the horizontal axis (i.e., the abscissa axis) of the rectangular coordinate system where the white blood cell histogram is located.

[0090] Furthermore, performing white blood cell classification and counting based on the second white blood cell histogram further includes: obtaining the classification results of each type of white blood cell according to the proportions of the first type of white blood cell, the second type of white blood cell, the third type of white blood cell, the fourth type of white blood cell, and the fifth type of white blood cell in the second white blood cell histogram respectively after classification. For example, obtaining the classification result of the first type of white blood cell as LYM2%, the classification result of the second type of white blood cell as MON2%, the classification result of the third type of white blood cell as NEU2%, the classification result of the fourth type of white blood cell as EOS2%, and the classification result of the fifth type of white blood cell as BASO2%. When it is four-class classification, it is as shown in Table 1 below, and when it is five-class classification, it is as shown in Table 2 below; calculating the count results of each type of white blood cell based on the classification results of each type of white blood cell and the count result of the white blood cell. For example, as shown in Table 1, the count result of the white blood cell in the second white blood cell histogram is WBC2, and this count result of the white blood cell is also the total number of white blood cell particles detected in the blood sample. It can be this count result directly obtained based on the number of pulse signals obtained when detecting the blood sample by the impedance method, or it can be calculated based on the produced second white blood cell histogram. For example, it can be obtained by adding up the particle numbers corresponding to the ordinates at each integer volume of the second white blood cell histogram and then taking the integer, or other counting methods.

[0091] Among them, calculating the count results of each type of white blood cell based on the classification results of each type of white blood cell and the count result of the white blood cell may include: multiplying the classification result of each type of white blood cell by the count result of the white blood cell and then calculating the count results of each type of white blood cell, as shown in Table 1 and Table 2 below.

[0092] Table 1 List of four-class classification results and count results

[0093]

[0094] The list of five-class classification results and count results is shown in Table 2 as follows:

[0095] Table 2 List of five-class classification results and count results

[0096]

[0097] Finally, as Figure 13 and Figure 14As shown in Table 1 and Table 2, the four-classification results and count results, and five-classification results and count results of the second white blood cell histogram are directly used as the four-classification results and count results, and five-classification results and count results of the white blood cells in the final blood sample. Since the blood shadow value in the second white blood cell histogram determined to be bimodal is less than the threshold blood shadow value, it is hardly interfered by the blood shadow. Therefore, its count result can accurately reflect the count result of white blood cells in the blood sample. At the same time, because its peak type is bimodal and each particle group in the white blood cells is relatively separated, it is conducive to the four-classification or five-classification of white blood cells and can obtain accurate classification results. Therefore, using the classification results and count results of the second white blood cell histogram determined to be bimodal as the classification results and count results of the final white blood cells has higher accuracy.

[0098] Further, as Figure 4 shown, in step S404, when the peak type of the second white blood cell histogram is unimodal, the white blood cell classification and counting are performed by combining the classification results and count results of the second white blood cell histogram and the classification results of the first white blood cell histogram.

[0099] In one example, the combining the classification and count results of the second white blood cell histogram and the classification results of the first white blood cell histogram for white blood cell classification and counting includes the following steps:

[0100] First, classify and count the second white blood cell histogram determined to be unimodal to obtain the count result of white blood cells and the classification result of eosinophils and / or basophils in the white blood cells.

[0101] In one example, only eosinophils (EOS) can be classified in the second white blood cell histogram determined to be unimodal. Eosinophils (EOS) can be classified by any suitable method. For example, as Figure 8 shown, the method for classifying the second white blood cell histogram determined to be unimodal includes: determining the peak point A of the second white blood cell histogram. Since the second white blood cell histogram is unimodal, this peak point A is the maximum value point on the second white blood cell histogram, that is, the point with the largest ordinate value. Then, starting from the peak point A, search along the direction of increasing volume for the first demarcation point B on the curve of the second white blood cell histogram where the slope is greater than the first threshold slope K eos for the first time. Among them, the region of the second white blood cell histogram with a volume greater than the volume of this first demarcation point B is the eosinophils.

[0102] The value of the first threshold slope K eos can be reasonably set according to actual needs. For example, the first threshold slope K eos is less than 0.

[0103] In another example, basophils (BASO) can also be classified only in the second white blood cell histogram determined to be unimodal. For example, basophils (BASO) can be classified in the second white blood cell histogram by the following method, including determining a fourth boundary line starting from the maximum volume Vmax (e.g., 250 fL) in the direction of decreasing volume. The fourth boundary line is spaced from the maximum volume by a predetermined volume (e.g., about 50 fL). This predetermined volume can be reasonably set according to prior experience and is not specifically limited here. Among them, the region on the second white blood cell histogram with a volume greater than the volume of the fourth boundary line is the basophil.

[0104] In other examples, as Figure 9 shown, eosinophils and basophils can also be classified in the second histogram determined to be unimodal. For example, the first demarcation point B can be determined by the method as Figure 8 shown, and then the fourth boundary line between the eosinophils and the basophils is determined according to the third boundary line passing through the first demarcation point B. Among them, the fourth boundary line is spaced from the third boundary line by a predetermined volume S baso , and this predetermined volume S baso can be reasonably set according to prior experience. For example, this predetermined volume S baso is about 75 fL, and the volume of the fourth boundary line is greater than the volume of the third boundary line. The region between the third boundary line and the fourth boundary line is the eosinophil EOS, the region on the second white blood cell histogram with a volume greater than the volume of the fourth boundary line is the basophil BASO, and the region between the fourth boundary line and the maximum volume.

[0105] The above method for classifying eosinophils and / or basophils in the second histogram determined to be unimodal is only an example, and other suitable methods can also be equally applicable to this application.

[0106] Optionally, the classification result of the eosinophil is the proportion of the region where the eosinophil is located in the second white blood cell histogram. For example, ESO2% shown in Table 1 and Table 2; the classification result of the basophil is the proportion of the region where the eosinophil is located in the second white blood cell histogram. For example, BASO2% shown in Table 2.

[0107] After the step of determining the peak shape of the second white blood cell histogram, or before the step of determining the peak shape, at least three classifications can be performed on the first white blood cell histogram. In this embodiment, mainly the case of performing three classifications on the first white blood cell histogram is taken as an example. For example, after the step of determining the peak shape of the second white blood cell histogram, if it is determined that the second white blood cell histogram is unimodal and the peak shape of the first white blood cell histogram is bimodal, at least three classifications are performed on the first white blood cell histogram.

[0108] In one example, the method for determining that the second white blood cell histogram is unimodal and performing three classifications on the first white blood cell histogram includes: demarcating a ghost cell region in the first white blood cell histogram; removing the ghost cell region from the first white blood cell histogram; and performing three classifications on the first white blood cell histogram after removing the ghost cell region.

[0109] The ghost cell region in the first white blood cell histogram can be demarcated by any suitable method. For example, the boundary line between the ghost cell and the white blood cell is found; the ghost cell region is demarcated according to the boundary line, where the region in the first white blood cell histogram with a volume smaller than the volume of the boundary line is the ghost cell region.

[0110] The boundary line between the ghost cell and the white blood cell can be determined by any suitable method. In one example, as Figure 10 shown, finding the boundary line between the ghost cell and the white blood cell includes: determining the first peak point A on the first white blood cell histogram starting from the minimum volume and along the direction of increasing volume; determining the boundary line according to the first peak point A, where the boundary line is spaced a predetermined volume from the first peak point A, and the volume of the boundary line is greater than the volume of the first peak point A. For example, the predetermined volume can be about 46 fL.

[0111] Another method for determining the ghost cell region can be as Figure 11 shown, setting a fixed volume V, which can be reasonably set according to the actual situation. For example, if V is equal to about 45 fL, then the region in the first white blood cell histogram with a volume smaller than the fixed volume V is the ghost cell region.

[0112] Furthermore, the first white blood cell histogram after removing the ghost cell region can be classified into three categories by any suitable method. For example, as Figure 12As shown in the figure, the three-classification of the first white blood cell histogram after removing the blood shadow area includes: obtaining the first valley point C in the first white blood cell histogram along the direction of increasing volume starting from the dividing line between the blood shadow area and the white blood cells. The first valley point C can be determined by any suitable method. For example, starting from the first valley point C, the ordinate values of a predetermined number of points continuously increase along the direction of decreasing volume, while starting from the first valley point C, the ordinate values of a predetermined number of points continuously decrease along the direction of increasing volume; based on the first valley point, a first dividing line between the first type of white blood cells and the second type of white blood cells is delimited, where the volume of the first type of white blood cells is smaller than the volume of the first valley point, and the volume of the second type of white blood cells is larger than the volume of the first valley point; according to the first dividing line, a second dividing line between the second type of white blood cells and the third type of white blood cells is determined, and the first dividing line and the second dividing line are spaced by a predetermined volume S mon , Smon is, for example, 28 fL or other suitable values. Among them, the area between the dividing line between the white blood cells and the blood shadow area and the first dividing line is the first type of white blood cells, the area between the first dividing line and the second dividing line is the second type of white blood cells, and the area with a volume larger than the volume of the second dividing line is the third type of white blood cells, that is, the area between the second dividing line and the maximum volume Vmax is the third type of white blood cells. Among them, the classification result of each type of white blood cell in the first white blood cell histogram is the proportion of each type of white blood cell in the first white blood cell histogram after removing the blood shadow area. Optionally, the first type of white blood cells is lymphocytes, the second type of white blood cells is monocytes, and the third type of white blood cells is neutrophils.

[0113] Optionally, the classification results of the first type of white blood cells, the second type of white blood cells, and the third type of white blood cells are the proportions of each type of white blood cell in the first white blood cell histogram after removing the blood shadow area. For example, as shown in Table 1 and Table 2, the first type of white blood cells is lymphocytes, and its classification result is LYM1%, the second type of white blood cells is monocytes MON, and its classification result is MON1%, and the third type of white blood cells is neutrophils, and its classification result is NEU1%.

[0114] Since the particle clusters in the second white blood cell histogram determined to be unimodal are closely spaced and it is difficult to perform more classifications, based on the classification results of the second white blood cell histogram and the classification results of the first white blood cell histogram that have been classified at least three times, where the first white blood cell histogram includes white blood cells of the first type, white blood cells of the second type, and white blood cells of the third type, at least a four-classification white blood cell classification result of the white blood cells in the final blood sample is obtained. For example, as shown in Table 1 and Table 2, the classification results of eosinophils and / or basophils in the second white blood cell histogram are used as the classification results of eosinophils and / or basophils of the white blood cells in the final blood sample; the classification result of the white blood cells of the third type (e.g., NEU1%) in the first white blood cell histogram that has been classified at least three times is subtracted by the classification result of eosinophils (ESO2%) and / or the classification result of basophils (BASO2%) in the second white blood cell histogram to obtain the classification result of the white blood cells of the third type (NEU%) in the final blood sample.

[0115] As Figure 13 In the example shown, when it is determined that the second white blood cell histogram is not bimodal, that is, the second white blood cell histogram is unimodal, after classifying eosinophils or basophils from the second white blood cell histogram determined to be unimodal, the classification results of eosinophils or basophils are combined with the three-classification results of the first white blood cell histogram after three-classification to obtain the four-classification result of the white blood cells in the final blood sample, which is shown in Table 1.

[0116] As Figure 14 In the example shown, when it is determined that the second white blood cell histogram is not bimodal, that is, the second white blood cell histogram is unimodal, after classifying eosinophils and basophils from the second white blood cell histogram determined to be unimodal, the classification results of eosinophils and basophils are combined with the three-classification results of the first white blood cell histogram after three-classification to obtain the five-classification result of the white blood cells in the final blood sample, which is shown in Table 2.

[0117] Since the interference of blood shadows in the second white blood cell histogram determined to be unimodal is almost negligible, the count result of white blood cells WBC2 in the second white blood cell histogram can be used as the count result of white blood cells in the final blood sample, that is, the result of the total number of white blood cells.

[0118] Furthermore, the count result of each type of white blood cell in the blood sample can also be calculated based on the at least four-class white blood cell classification result and the count result of the white blood cells. For example, if the four-classification result of the white blood cells in the final blood sample is shown in Table 1, then the count result LYM# of the first type of white blood cell, lymphocytes, is the product of the classification result LYM1% of lymphocytes in the first white blood cell histogram and the count result WBC2 of the white blood cells in the second white blood cell histogram. The count result MON# of the second type of white blood cell, monocytes, is the product of the classification result MON1% of monocytes in the first white blood cell histogram and the count result WBC2 of the white blood cells in the second white blood cell histogram. Finally, the count result of the third type of white blood cell in the first white blood cell histogram after at least three classifications (that is, the count result obtained by multiplying the classification result of the third type of white blood cell by the count result of the white blood cells in the second white blood cell histogram determined to be unimodal) minus the eosinophil count result in the second white blood cell histogram is used as the count result of the third type of white blood cell in the blood sample. Alternatively, it can also be that the classification result of the third type of white blood cell in the first white blood cell histogram after at least three classifications minus the classification result of eosinophils in the second white blood cell histogram is used as the classification result NEU% of the third type of white blood cell in the final blood sample. Then, multiplying this final classification result NEU% of the third type of white blood cell by the count result WBC2 of the white blood cells to obtain the final count result NEU# of the third type of white blood cell, and the eosinophil count result EOS# is the product of the classification result EOS2% of eosinophils in the second white blood cell histogram and the count result WBC2 of the white blood cells in the second white blood cell histogram.

[0119] For example, if the five-classification results of white blood cells in the final blood sample are shown in Table 2, then the counting result LYM# of lymphocytes, the first type of white blood cells, is the product of the classification result LYM1% of lymphocytes in the first white blood cell histogram and the counting result WBC2 of white blood cells in the second white blood cell histogram. The counting result MON# of monocytes, the second type of white blood cells, is the product of the classification result MON1% of monocytes in the first white blood cell histogram and the counting result WBC2 of white blood cells in the second white blood cell histogram. Finally, subtract the counting result of eosinophils and the counting result of basophils in the second white blood cell histogram from the counting result of the third type of white blood cells in the first white blood cell histogram that has been at least three-classified (that is, the counting result obtained by multiplying the classification result of the third type of white blood cells by the counting result of white blood cells in the second white blood cell histogram determined to be unimodal) as the counting result of the third type of white blood cells in the blood sample. Or, it can also be that subtract the classification result of eosinophils and the classification result of basophils in the second white blood cell histogram from the classification result of the third type of white blood cells in the first white blood cell histogram that has been at least three-classified as the classification result NEU% of the third type of white blood cells in the final blood sample, and then multiply the final classification result NEU% of the third type of white blood cells by the counting result WBC2 of white blood cells to obtain the counting result NEU# of the third type of white blood cells. The counting result EOS# of eosinophils is the product of the classification result EOS2% of eosinophils in the second white blood cell histogram and the counting result WBC2 of white blood cells in the second white blood cell histogram. The counting result BASO# of basophils is the product of the classification result BASO% of basophils in the second white blood cell histogram and the counting result WBC2 of white blood cells in the second white blood cell histogram.

[0120] Finally, use the classification results of the first type of white blood cells and the second type of white blood cells in the first white blood cell histogram that has been at least three-classified as the classification results of the first type of white blood cells and the second type of white blood cells in the blood sample. Finally, use the counting results of the first type of white blood cells and the second type of white blood cells in the first white blood cell histogram that has been at least three-classified as the counting results of the first type of white blood cells and the second type of white blood cells in the blood sample.

[0121] As Figure 15 and Figure 16 shown in the specific example, in Figure 15In the second white blood cell histogram obtained from the blood sample treated with the second hemolytic agent as shown, it is determined that the second white blood cell histogram is unimodal. At this time, the white blood cell count result WBC2 is 10.6. According to the method described above, the eosinophil Eos% is obtained as 7.1%, and for Figure 16 the first white blood cell histogram as shown is classified into three categories, and the classification results are shown in Table 3 below:

[0122] Table 3

[0123]

[0124] Then, according to the method of the foregoing embodiment and the rules in Table 1, the white blood cell count result and classification result of the blood sample can be finally obtained, as shown in Table 4:

[0125] Table 4

[0126]

[0127] As can be seen from Table 4, when the peak type of the second white blood cell histogram is unimodal, the count result and classification result obtained by the classification and counting method combining the second white blood cell histogram and the first white blood cell histogram are very close to the reference value (and the true value). Thus, it can be seen that it can significantly improve the accuracy of white blood cell counting and classification.

[0128] In summary, when the peak type of the second white blood cell histogram is unimodal, the white blood cell classification and counting are performed by combining the classification result and count result of the second white blood cell histogram and the classification result of the first white blood cell histogram. Since the distribution of white blood cell particle clusters is compact when the peak type of the second white blood cell histogram is unimodal, it is not conducive to classification, but its influence by the blood shadow is small. Therefore, its count result is more accurate than that of the first white blood cell histogram. And since the first white blood cell histogram is bimodal and the distribution of its white blood cell particle clusters is relatively dispersed, it is conducive to classification. Therefore, by combining the classification result and count result of the second white blood cell histogram determined to be unimodal and the classification result of the first white blood cell histogram for white blood cell classification and counting, a more accurate white blood cell classification and counting result can be obtained. Therefore, the method of the embodiment of the present invention can improve the accuracy of white blood cell counting and classification.

[0129] It can be understood that when performing four-classification of white blood cells in the present application, usually on the basis of three-classification, neutrophils are separated from eosinophils or basophils. For example, when white blood cells are divided into lymphocytes, monocytes, neutrophils, and eosinophils, at this time, the number of basophils is included in eosinophils; when white blood cells are divided into lymphocytes, monocytes, neutrophils, and basophils, the number of eosinophils is included in neutrophils. When generating a report, it can be displayed as the number of neutrophils, the total number of neutrophils and eosinophils, or the total number of granulocytes. That is to say, in this case, neutrophils and eosinophils are not counted separately.

[0130] In another embodiment, as Figure 17 shown, the method for classifying and counting white blood cells according to the embodiment of the present invention includes the following steps S501 to S504: Step S501, obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a hemolytic agent for a first treatment time and a second white blood cell histogram of white blood cells in a blood sample treated with the hemolytic agent for a second treatment time, wherein the second treatment time is greater than the first treatment time, the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value;

[0131] In step S501, a first white blood cell histogram of white blood cells in a blood sample treated with a hemolytic agent for a first treatment time and a second white blood cell histogram of white blood cells in a blood sample treated with the hemolytic agent for a second treatment time are obtained, wherein the second treatment time is greater than the first treatment time.

[0132] The first treatment time and the second treatment time can be reasonably set according to actual reaction conditions, etc. For example, the first treatment time can be zero or a treatment time greater than 0 set according to prior experience. This first treatment time needs to ensure that there is a blood shadow in the first white blood cell histogram of white blood cells in the blood sample treated with the hemolytic agent for the first treatment time, and the blood shadow value is greater than or equal to the threshold blood shadow. The existence of this blood shadow will affect the accuracy of the white blood cell counting result, and the obtained first white blood cell histogram is bimodal. The second treatment time can be to continue to increase the treatment time after the first treatment time with the hemolytic agent until the predetermined second treatment time is reached, and then generate the second white blood cell histogram. At this time, the blood shadow in the blood sample is significantly reduced or even disappears compared with the blood sample treated with the first treatment time. This second treatment time needs to ensure that the blood shadow value in the obtained second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than the threshold blood shadow value.

[0133] Among them, in this embodiment, it is possible to ensure that a sufficient amount of hemolytic agent is added. The amount of the hemolytic agent needs to meet the requirement that there is less blood shadow in the processed blood sample at the second processing time, so that the generated second white blood cell histogram meets the following requirements: the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than the threshold blood shadow value.

[0134] Since the hemolytic agent takes a certain processing time to dissolve all the red blood cells, and at the shorter first processing time, only part of the red blood cells are dissolved and there are more blood shadows in the blood sample at this time, so the first white blood cell histogram can be obtained. As the processing time increases and reaches the second processing time, more red blood cells are dissolved and the blood shadows in the blood sample decrease, so the second white blood cell histogram can be obtained.

[0135] Subsequently, in step S502, determine the peak shape of the second white blood cell histogram; in step S503, when the peak shape of the second white blood cell histogram is bimodal, perform white blood cell classification and counting with the second white blood cell histogram; in step S504, when the peak shape of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram.

[0136] Among them, the descriptions of steps S502 to S504 in this embodiment can refer to the descriptions of steps S402 to S404 in the foregoing embodiment, and will not be repeated here to avoid redundancy. Through the method of the embodiment of the present invention, only one hemolytic agent needs to be added, and by adjusting the processing time, the first white blood cell histogram and the second white blood cell histogram can be obtained. Thus, when the peak shape of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram. Since the distribution of white blood cell particle clusters is compact when the peak shape of the second white blood cell histogram is unimodal, it is not conducive to classification, but it is less affected by blood shadows, so its counting result is more accurate than that of the first white blood cell histogram. And since the first white blood cell histogram is bimodal and the distribution of its white blood cell particle clusters is relatively dispersed, which is conducive to classification. Therefore, by combining the classification result and counting result of the second white blood cell histogram determined to be unimodal and the classification result of the first white blood cell histogram for white blood cell classification and counting, more accurate white blood cell classification and counting results can be obtained. When the second white blood cell histogram is bimodal, directly perform white blood cell classification and counting with the second white blood cell histogram, and accurate white blood cell counting and classification results can also be obtained. Therefore, the method of the embodiment of the present invention can improve the accuracy of white blood cell counting and classification.

[0137] Next, refer toFigure 18 An example electronic device 150 for implementing the method of white blood cell classification and counting according to an embodiment of the present invention is described.

[0138] In one example, as Figure 18 shown, the electronic device 150 may include one or more processors 151, one or more storage devices 152, an input device 153, an output device 154, and a communication interface 155, and these components are interconnected through a bus system 156 and / or other forms of connection mechanisms (not shown). It should be noted that Figure 18 the components and structures of the electronic device 150 shown are exemplary and not restrictive. According to needs, the electronic device may also have other components and structures.

[0139] The processor 151 may be a central processing unit (CPU), an image processing unit (GPU), an application specific integrated circuit (ASIC), a field programmable gate array (FPGA), or other forms of processing units having data processing capabilities and / or instruction execution capabilities, and may control other components in the white blood cell classification and counting device to perform desired functions. The processor is capable of executing programs and / or instructions stored in the storage device to perform the method of white blood cell classification and counting described herein. For example, the processor 151 may include one or more embedded processors, processor cores, microprocessors, logic circuits, hardware finite state machines (FSMs), digital signal processors (DSPs), or combinations thereof.

[0140] The storage device 152 may include one or more computer program products, and the computer program products may include various forms of computer-readable storage media, such as volatile memory and / or non-volatile memory. The volatile memory may include, for example, random access memory (RAM) and / or cache memory, etc. The non-volatile memory may include, for example, read-only memory (ROM), hard disk, flash memory, etc. One or more computer program instructions may be stored on the computer-readable storage media, and the processor 151 may run the program instructions to implement the functions (implemented by the processor) in the embodiments of the present invention described herein and / or other desired functions. Various application programs and various data may also be stored in the computer-readable storage media, such as various data used and / or generated by the application programs, etc.

[0141] The input device 153 can be a device used by a user to input instructions (for example, in the method of white blood cell classification and counting according to an embodiment of the present invention described herein, the user can input parameters such as a predetermined volume when dividing a boundary line), and can include one or more of a keyboard, a mouse, a microphone, a touch screen, and the like. In addition, the input device 153 can also be any interface for receiving information.

[0142] The output device 154 can output various information (such as images or sounds) to the outside (such as a user), and can include one or more of a display (such as displaying a parameter list of a white blood cell classification and counting device, the result of white blood cell classification and counting, and a histogram of white blood cell classification to the user), a speaker, and the like.

[0143] The communication interface 155 is used for communication between the electronic device 150 and other devices, including wired or wireless communication. The electronic device 150 can access a wireless network based on a communication standard, such as WiFi, 2G, 3G, 4G, 5G, or a combination thereof. In an exemplary embodiment, the communication interface 155 further includes a near field communication (NFC) module to facilitate short-range communication. For example, the NFC module can be implemented based on radio frequency identification (RFID) technology, infrared data association (IrDA) technology, ultra-wideband (UWB) technology, Bluetooth (BT) technology, and other technologies.

[0144] Exemplarily, an example electronic device for implementing the method of white blood cell classification and counting according to an embodiment of the present invention can be implemented as a terminal such as a desktop computer, a tablet computer, or the like, or a blood analyzer including these terminals.

[0145] In addition, an embodiment of the present invention further provides a computer-readable storage medium, on which a computer program is stored. One or more computer program instructions can be stored on the computer-readable storage medium, and a processor can run the program instructions stored in the storage device to implement the functions (implemented by the processor) in the embodiments of the present invention described herein and / or other desired functions, such as performing corresponding steps of the method of white blood cell classification and counting according to an embodiment of the present invention. Various application programs and various data can also be stored in the computer-readable storage medium, such as various data used and / or generated by the application programs.

[0146] For example, the computer-readable storage medium can include, for example, a memory card of a smart phone, a storage component of a tablet computer, a hard disk of a personal computer, a read-only memory (ROM), an erasable programmable read-only memory (EPROM), a portable compact disc read-only memory (CD-ROM), a USB memory, or any combination of the above storage media.

[0147] Next, in conjunction withFigure 19 Describe the blood sample analyzer provided by another aspect of the present invention. Figure 19 Show a schematic block diagram of a blood analyzer in an embodiment of the present invention.

[0148] The blood analyzer is used for various analyses of blood components, such as counting and classifying white blood cells in blood, detecting the concentration of hemoglobin (HGB) in red blood cells, counting platelets, etc.

[0149] As Figure 19 Shown, the blood sample analyzer includes at least one reaction cell 182 and a sampling needle assembly 181. The reaction cell 182 is used to prepare a blood sample from the blood sample to be tested, and the sampling needle assembly 181 is used to discharge the blood sample to be analyzed into the reaction cell 182. In another embodiment, the sampling needle assembly 181 can also be implemented in other ways other than the sampling needle.

[0150] In one example, the blood analyzer further includes a reagent storage device (not shown), which is connected to the reaction cell 182 and is used to provide reagents for preparing blood samples to the reaction cell 182, such as hemolytic agents, diluents, etc. Among them, the number of reagent storage devices is reasonably set according to the types of reagents. For example, the reagent storage device includes a storage device for storing hemolytic agents and a storage device for storing diluents. Among them, the storage device for storing hemolytic agents can also be divided into one or more according to the types of hemolytic agents. For example, a storage device for storing the first hemolytic agent and a storage device for storing the second hemolytic agent. When the first hemolytic agent and the second hemolytic agent are the same hemolytic agent, only one hemolytic agent storage device can be set.

[0151] In one example, the blood analyzer 180 further includes a hemolytic agent pushing component 183, which is used to push the hemolytic agent into the reaction cell 182. The hemolytic agent includes the first hemolytic agent and the second hemolytic agent. Exemplarily, the hemolytic agent pushing component 183 can be connected to the hemolytic agent storage device and the reaction cell, so as to push the corresponding hemolytic agent into the reaction cell 182. After injecting the reagent into the reaction device, the blood sample, hemolytic agent, diluent, etc. are mixed in the reaction cell 182 to prepare a blood sample for detecting white blood cells.

[0152] The blood analyzer 180 further includes a conveying device (not shown), which is used to convey the sample liquid in the reaction cell 182 to the resistive detector 184. In this embodiment, the conveying device includes a syringe and a conveying pipeline communicated with the syringe. The syringe, the sample liquid output port of the reaction cell and the entrance of the detection device are communicated through the conveying pipeline. There can be multiple syringes, and each syringe performs suction and discharge actions under the control of the control device.

[0153] In one example, as Figure 19As shown, the blood analyzer 180 further includes a resistive detector 184. The resistive detector 184 is configured to detect white blood cells in the blood sample treated with the first hemolytic agent and generate a first signal, such as a pulse signal, and detect white blood cells in the blood sample treated with the second hemolytic agent and generate a second signal, such as a pulse signal. For example, the resistive detector 184 is configured to detect white blood cells in the blood sample and output a pulse signal when the white blood cells pass through the detection aperture in the resistive detector 184. Among them, the number of the pulse signals is proportional to the number of cells, and the height of the pulse signal is proportional to the cell volume, thereby obtaining the number and volume values of blood cells in the blood. Among them, the resistive detector 184 is configured to detect the blood sample prepared by the reaction cell.

[0154] In one example, the structure of the resistive detector 184 may include a counting cell (not shown) and a pulse sensor (not shown). Among them, the counting cell and the reaction cell may be the same device or different devices. Optionally, the pulse sensor may include a small hole tube, and a detection aperture is provided on the small hole tube. Optionally, the diameter of the detection aperture is less than 100 microns, and the thickness ranges from 60 microns to 90 microns, for example, about 75 microns. The counting cell is filled with a conductive solvent and is divided into a front cell and a rear cell by the detection aperture; a positive electrode and a negative electrode are respectively provided in the front cell and the rear cell. The positive and negative electrodes are connected to one end of a constant current source. The electrode provided in the front cell, the conductive solvent, the electrode provided in the rear cell, and the constant current source together form a series closed loop. When the power is turned on, a stable current is generated between the electrodes on both sides of the small hole tube. The diluted cell suspension flows from the outside of the small hole tube through the detection aperture into the inside of the small hole tube, increasing the resistance in the small hole induction area and causing an instantaneous voltage change to form a pulse signal. The amplitude of the pulse signal is proportional to the cell volume, and the number of pulses is proportional to the number of cells, thereby obtaining the number and volume values of blood cells in the blood sample, and different types of cells can be distinguished according to the volume distribution.

[0155] In another embodiment, the sampling needle assembly is configured to discharge the blood sample to be analyzed into the reaction cell; the hemolytic agent pushing component is configured to push the hemolytic agent into the reaction cell; the resistive detector is configured to detect white blood cells in the blood sample treated with the hemolytic agent for a first treatment time and generate a first signal, and detect white blood cells in the blood sample treated with the hemolytic agent for a second treatment time and generate a second signal, where the second treatment time is greater than the first treatment time.

[0156] In one example, the hematology analyzer 180 further includes one or more processors 185, which work jointly or individually to obtain a first white blood cell histogram based on the first signal and a second white blood cell histogram based on the second signal, wherein the ghost cell value in the second white blood cell histogram is less than the ghost cell value in the first white blood cell histogram, and the ghost cell value in the second white blood cell histogram is less than a threshold ghost cell value. For example, during each detection, the processor 185 receives signals output by the resistive detector 184, such as the first signal and the second signal, and analyzes the first signal and the second signal, so as to obtain a first white blood cell histogram and a second white blood cell histogram based on the first signal and the second signal.

[0157] In one embodiment, the hematology analyzer 180 includes one or more processors 185 and one or more storage devices (not shown). The one or more processors work jointly or individually, and the storage device stores a program for implementing corresponding steps in the method for white blood cell classification and counting according to the embodiments of the present invention. The processor is configured to run the program stored in the storage device to execute the corresponding steps of the method for white blood cell classification and counting according to the embodiments of the present invention.

[0158] In one embodiment of the present invention, when the program is run by the processor, the hematology analyzer 180 is caused to perform the following steps:

[0159] Obtain a first white blood cell histogram based on the first signal and a second white blood cell histogram based on the second signal, wherein the ghost cell value in the second white blood cell histogram is less than the ghost cell value in the first white blood cell histogram, and the ghost cell value in the second white blood cell histogram is less than a threshold ghost cell value;

[0160] When the peak pattern of the second white blood cell histogram is a bimodal peak, perform white blood cell classification and counting using the second white blood cell histogram;

[0161] When the peak pattern of the second white blood cell histogram is a unimodal peak, perform white blood cell classification and counting by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram.

[0162] In one example, the sampling needle assembly is configured to discharge the blood sample to be analyzed into the reaction cell for the first time for the first measurement and discharge the blood sample to be analyzed into the reaction cell for the second time for the second measurement; the lysing agent pushing member is configured to: push the first lysing agent into the reaction cell to complete the first measurement and generate the first white blood cell histogram; push the second lysing agent into the reaction cell to complete the second measurement and generate the second white blood cell histogram.

[0163] In another example, the sampling needle assembly 181 is used to discharge a blood sample to be analyzed into the reaction cell 182. The hemolytic agent pushing member is used to: first push the first hemolytic agent into the blood sample in the reaction cell to obtain a test solution, so as to complete the first measurement and obtain the first white blood cell histogram; then push the second hemolytic agent into the test solution in the reaction cell to complete the second measurement and obtain the second white blood cell histogram.

[0164] In other examples, the hemolytic agent pushing member is used to: sequentially push at least three hemolytic agents into the same blood sample in the reaction cell, and each added hemolytic agent includes at least one of the first hemolytic agent and the second hemolytic agent; the resistive detector is used to: detect white blood cells in the blood sample after each time the hemolytic agent pushing member pushes the first hemolytic agent and / or the second hemolytic agent to generate corresponding signals; the processor is used to: generate corresponding white blood cell histograms based on the corresponding signals, and select two successively generated white blood cell histograms in the corresponding white blood cell histograms as the first white blood cell histogram and the second white blood cell histogram respectively.

[0165] In one embodiment, when the program is run by the processor, the blood analyzer 180 is caused to perform the following steps: classify and count the second white blood cell histogram determined to be unimodal to obtain a white blood cell count result and a classification result of eosinophils and / or basophils in the white blood cells; obtain a classification result of at least four classifications according to the classification result of the second white blood cell histogram and the classification result of the first white blood cell histogram that has been at least three-classified; calculate the count result of each type of white blood cell in the blood sample based on the classification result of at least four classifications and the white blood cell count result.

[0166] The first white blood cell histogram that has been at least three-classified includes a first type of white blood cell, a second type of white blood cell, and a third type of white blood cell. In one embodiment, when the program is run by the processor, the blood analyzer 180 is caused to perform the following steps: use the count result of eosinophils and / or basophils in the second white blood cell histogram as the count result of eosinophils and / or basophils in the blood sample; subtract the count result of eosinophils and / or basophils in the second white blood cell histogram from the count result of the third type of white blood cell in the first white blood cell histogram that has been at least three-classified as the count result of the third type of white blood cell in the blood sample; use the count result of the first type of white blood cell and the count result of the second type of white blood cell in the first white blood cell histogram that has been at least three-classified as the count result of the first type of white blood cell and the second type of white blood cell in the blood sample.

[0167] In one embodiment, the classification result of each type of white blood cells in the first white blood cell histogram is the proportion of each type of white blood cells in the first white blood cell histogram after removing the blood ghost area; and / or the classification result of the eosinophils is the proportion of the area where the eosinophils are located in the second white blood cell histogram; the classification result of the basophils is the proportion of the eosinophils in the second white blood cell histogram.

[0168] In one embodiment, the blood analyzer further includes a detection mode selection module for selecting a predetermined detection mode according to the animal type to which the blood sample belongs; the hemolytic agent pushing component pushes the corresponding hemolytic agent to the reaction pool for reaction according to the selected detection mode.

[0169] In one embodiment, when the program is executed by the processor, the blood analyzer 180 executes the relevant steps of the method for classifying and counting white blood cells described in the above embodiments.

[0170] A complete blood analyzer may also include other components, which will not be described in detail here.

[0171] In summary, according to the method for classifying and counting white blood cells, the blood analyzer, and the storage medium of the embodiment of the present invention, a first white blood cell histogram and a second white blood cell histogram of white blood cells in a blood sample treated with a first hemolytic agent and a second hemolytic agent are obtained, and the white blood cell count and classification method is determined according to the peak type of the second white blood cell histogram. When the peak type of the second white blood cell histogram is bimodal and the blood ghost value in the second white blood cell histogram is less than the threshold blood ghost value, the blood ghost will hardly interfere with the counting of the second white blood cell histogram. Moreover, since it is bimodal, the regional distribution of various types of white blood cell particle clusters in the second white blood cell histogram is relatively open. Therefore, the classification and counting results of white blood cells in the blood can be accurately obtained by directly counting with the second white blood cell histogram. When the peak type of the second white blood cell histogram is unimodal, The classification and counting results of the second white blood cell histogram are combined with the classification results of the first white blood cell histogram to perform white blood cell classification and counting. Since the second white blood cell histogram has a single peak, the distribution of its white blood cell clusters is compact, making classification difficult, but it is less affected by blood shadows, so its count result is more accurate than that of the first white blood cell histogram. Since the first white blood cell histogram is bimodal, the distribution of its white blood cell clusters is more open, making classification easier. Therefore, by combining the classification and counting results of the second white blood cell histogram, which is determined to be single-peaked, with the classification results of the first white blood cell histogram, a more accurate white blood cell classification and counting result can be obtained. Therefore, the method of the embodiment of the present invention can improve the accuracy of white blood cell counting and classification. Thus, the counting and classification results more accurately reflect the actual condition of the blood sample, allowing doctors and others to use these results to reasonably judge the health status of the blood sample source and make reasonable medical diagnoses.

[0172] Although example embodiments have been described herein with reference to the accompanying drawings, it should be understood that the above example embodiments are merely illustrative and are not intended to limit the scope of the present invention. Various changes and modifications may be made therein by those skilled in the art without departing from the scope and spirit of the present invention. All such changes and modifications are intended to be included within the scope of the present invention as claimed in the appended claims.

[0173] Those skilled in the art will appreciate that the units and algorithm steps of each example described in conjunction with the embodiments disclosed herein can be implemented in electronic hardware, or a combination of computer software and electronic hardware. Whether these functions are performed in hardware or software depends on the specific application and design constraints of the technical solution. Professionals and technicians can use different methods to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of the present invention.

[0174] In the several embodiments provided in this application, it should be understood that the disclosed devices and methods can be implemented in other ways. For example, the device embodiments described above are merely illustrative. For example, the division of the units described is merely a logical function division. In actual implementation, other division methods may be used, such as combining or integrating multiple units or components into another device, or ignoring or not performing some features.

[0175] In the description provided herein, a large number of specific details are described. However, it is understood that embodiments of the present invention can be practiced without these specific details. In some instances, well-known methods, structures and techniques are not shown in detail so as not to obscure the understanding of this description.

[0176] Similarly, it should be understood that in order to streamline the present invention and aid in understanding one or more of the various inventive aspects, in the description of exemplary embodiments of the present invention, the various features of the present invention are sometimes grouped together into a single embodiment, figure, or description thereof. However, this approach to the present invention should not be interpreted as reflecting the intention that the claimed invention requires more features than those explicitly recited in each claim. More precisely, as reflected in the corresponding claims, the inventive point is that the corresponding technical problem can be solved with fewer features than all the features of a single disclosed embodiment. Therefore, the claims following the detailed description are hereby expressly incorporated into the detailed description, with each claim itself serving as a separate embodiment of the present invention.

[0177] Those skilled in the art will understand that, except where mutually exclusive, all features disclosed in this specification (including the accompanying claims, abstract, and drawings) and all processes or units of any method or apparatus disclosed herein may be combined in any combination. Unless expressly stated otherwise, each feature disclosed in this specification (including the accompanying claims, abstract, and drawings) may be replaced by an alternative feature that provides the same, equivalent, or similar purpose.

[0178] Furthermore, those skilled in the art will appreciate that although some embodiments described herein include certain features included in other embodiments but not other features, combinations of features from different embodiments are intended to be within the scope of the present invention and to form different embodiments. For example, in the claims, any of the claimed embodiments may be used in any combination.

[0179] Each component embodiment of the present invention may be implemented in hardware, or in software modules running on one or more processors, or in a combination thereof. Those skilled in the art should understand that a microprocessor or a digital signal processor (DSP) may be used in practice to implement some or all of the functions of some modules according to the embodiments of the present invention. The present invention may also be implemented as a device program (e.g., a computer program and a computer program product) for performing part or all of the methods described herein. Such a program for implementing the present invention may be stored on a computer-readable medium, or may be in the form of one or more signals. Such signals may be downloaded from an Internet website, or provided on a carrier signal, or in any other form.

[0180] It should be noted that the above embodiments illustrate the present invention rather than limit the present invention, and those skilled in the art can design alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim. The present invention can be implemented by means of hardware including several different elements and by means of a suitably programmed computer. In the unit claims listing several devices, several of these devices may be embodied by the same hardware item. The use of the words first, second, and third, etc. does not denote any order. These words may be interpreted as names.

Claims

1. A method for white blood cell classification and counting, characterized in that, The method includes: Obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second lysing agent, wherein the ghost cell value in the second white blood cell histogram is less than the ghost cell value in the first white blood cell histogram, and the ghost cell value in the second white blood cell histogram is less than a threshold ghost cell value; Determining the peak type of the second white blood cell histogram; When the peak type of the second white blood cell histogram is bimodal, performing white blood cell classification and counting using the second white blood cell histogram; When the peak type of the second white blood cell histogram is unimodal, performing white blood cell classification and counting by combining the classification result and counting result of the second white blood cell histogram and the classification result of the first white blood cell histogram.

2. The method according to claim 1, characterized in that, The first lysing agent and the second lysing agent are the same lysing agent.

3. The method according to claim 1, characterized in that, The dose of the second lysing agent is greater than the dose of the first lysing agent.

4. The method according to any one of claims 1 to 3, characterized in that, The method further includes: First adding the first lysing agent to the blood sample to obtain a test solution, and obtaining the first white blood cell histogram; Then adding the second lysing agent to the test solution to obtain the second white blood cell histogram.

5. The method according to any one of claims 1 to 3, characterized in that Obtaining a first white blood cell histogram of white blood cells in a blood sample treated with a first lysing agent and a second white blood cell histogram of white blood cells in a blood sample treated with a second lysing agent includes: Sequentially adding at least three times of lysing agent to the same blood sample, each addition of the lysing agent including at least one of the first lysing agent and the second lysing agent, and generating a corresponding white blood cell histogram after each addition of the first lysing agent and / or the second lysing agent; Selecting two successively generated white blood cell histograms as the first white blood cell histogram and the second white blood cell histogram respectively.

6. The method according to claim 1, wherein The combining the classification and counting results of the second white blood cell histogram and the classification result of the first white blood cell histogram to perform white blood cell classification and counting includes: Classifying and counting the second white blood cell histogram determined to be unimodal to obtain a counting result of white blood cells and a classification result of eosinophils and / or basophils in the white blood cells; According to the classification result of the second white blood cell histogram and the classification result of the first white blood cell histogram that has been at least three-classified, the first white blood cell histogram includes a first type of white blood cell, a second type of white blood cell, and a third type of white blood cell, obtaining a white blood cell classification result of at least four classes; Calculating the counting result of each type of white blood cell in the blood sample based on the white blood cell classification result of at least four classes and the counting result of the white blood cells.

7. The method according to claim 6, wherein The calculating the counting result of each type of white blood cell in the blood sample based on the white blood cell classification result of at least four classes and the counting result of the white blood cells includes: Taking the counting result of eosinophils and / or basophils in the second white blood cell histogram as the counting result of eosinophils and / or basophils in the blood sample; Subtract the count result of the third type of white blood cells in the first white blood cell histogram after at least three classifications from the count result of eosinophils and / or basophils in the second white blood cell histogram as the count result of the third type of white blood cells in the blood sample; Use the count results of the first type of white blood cells and the second type of white blood cells in the first white blood cell histogram after at least three classifications as the count results of the first type of white blood cells and the second type of white blood cells in the blood sample.

8. The method according to claim 6 or 7, wherein The classification results of the first type of white blood cells, the second type of white blood cells, and the third type of white blood cells are the proportions of each type of white blood cells in the first white blood cell histogram after removing the blood shadow area; The classification result of eosinophils is the proportion of the area where eosinophils are located in the second white blood cell histogram; and / or The classification result of basophils is the proportion of the area where eosinophils are located in the second white blood cell histogram.

9. The method according to any one of claims 1-3, 6, and 7, characterized in that The method further includes: After the step of determining the peak type of the second white blood cell histogram, if it is determined that the second white blood cell histogram is unimodal and the peak type of the first white blood cell histogram is bimodal, perform at least three classifications on the first white blood cell histogram.

10. The method according to any one of claims 1-3, 6, and 7, characterized in that, Determining that the second white blood cell histogram is unimodal, and the method for performing three classifications on the first white blood cell histogram includes: Define a blood shadow area in the first white blood cell histogram; Remove the blood shadow area in the first white blood cell histogram; Perform three classifications on the first white blood cell histogram after removing the blood shadow area.

11. The method according to any one of claims 1 - 3, 6, and 7, characterized in that, The method for classifying the determined unimodal second white blood cell histogram includes: Determine the peak point of the second white blood cell histogram; Starting from the peak point, find the first demarcation point on the curve of the second white blood cell histogram where the slope is greater than the first threshold slope for the first time in the direction of increasing volume. Among them, the area of the second white blood cell histogram with a volume greater than the volume of this first demarcation point is eosinophils.

12. The method according to claim 11, wherein The method for classifying the determined unimodal second white blood cell histogram further includes: Determine the fourth demarcation line between eosinophils and basophils according to the third demarcation line passing through the first demarcation point. Among them, the fourth demarcation line is separated from the third demarcation line by a predetermined volume, and the volume of the fourth demarcation line is greater than the volume of the third demarcation line. The area between the third demarcation line and the fourth demarcation line is the eosinophils, and the area of the second white blood cell histogram with a volume greater than the volume of the fourth demarcation line is the basophils.

13. The method according to claim 11, wherein The first threshold slope is less than 0.

14. The method according to any one of claims 1-3, 6, and 7, characterized in that The method for classifying the determined unimodal second white blood cell histogram includes: Determine the fourth demarcation line starting from the maximum volume in the direction of decreasing volume. The fourth demarcation line is separated from the maximum volume by a predetermined volume. Among them, the area of the second white blood cell histogram with a volume greater than the volume of the fourth demarcation line is basophils.

15. The method according to claim 1, wherein The method includes: Determine the peak pattern of the second white blood cell histogram according to the number of peak points in the second white blood cell histogram. Among them, if the number of peak points is 2 and the distance between the two peak points is greater than a preset distance, the second white blood cell histogram is bimodal; otherwise, the second white blood cell histogram is unimodal.

16. The method according to any one of claims 1 to 3, characterized in that, A method for four-classifying the second white blood cell histogram determined to be bimodal includes: Determine the first boundary line between the first type of white blood cells and the second type of white blood cells according to the trough point between the two peak points in the second white blood cell histogram, where the volume of the first type of white blood cells is smaller than the volume of the second type of white blood cells. Determine the second boundary line between the second type of white blood cells and the third type of white blood cells according to the first boundary line, where the second boundary line and the first boundary line are separated by a first predetermined volume, and the volume corresponding to the second boundary line is greater than the volume corresponding to the first boundary line. Start from the maximum volume of the second white blood cell histogram and search along the direction of decreasing volume for the second critical point on the curve of the second white blood cell histogram where the slope is greater than the second threshold slope for the first time. Determine the third boundary line between the third type of white blood cells and the fourth type of white blood cells based on the second critical point.

17. The method according to claim 16, wherein A method for five-classifying the second white blood cell histogram determined to be bimodal includes: Determine the fourth boundary line between the fourth type of white blood cells and the fifth type of white blood cells according to the third boundary line, where the fourth boundary line and the third boundary line are separated by a second predetermined volume, and the volume corresponding to the fourth boundary line is greater than the volume corresponding to the third boundary line. The fifth type of white blood cells is the area on the second white blood cell histogram with a volume greater than the volume corresponding to the fourth boundary line.

18. The method according to claim 17, wherein The first type of white blood cells is lymphocytes, the second type of white blood cells is monocytes, the third type of white blood cells is neutrophils, the fourth type of white blood cells is eosinophils, and the fifth type of white blood cells is basophils.

19. The method according to claim 17, wherein Using the second white blood cell histogram for white blood cell classification and counting further includes: Obtain the classification results of each type of white blood cell according to the proportions of the first type of white blood cells, the second type of white blood cells, the third type of white blood cells, the fourth type of white blood cells, and the fifth type of white blood cells in the second white blood cell histogram after classification. Calculate the count results of each type of white blood cell according to the classification results of each type of white blood cell and the count results of the white blood cells.

20. The method according to any one of claims 1 to 3, 6, 7, 12, 13, 15 and 17 - 19, characterized in that, Both the first white blood cell histogram and the second white blood cell histogram are obtained based on impedance measurement of a blood sample.

21. The method according to any one of claims 1 to 3, 6, 7, 12, 13, 15 and 17 - 19, characterized in that, The blood sample is an animal blood sample.

22. The method according to any one of claims 1 to 3, 6, 7, 12, 13, 15 and 17-19, characterized in that, The method further includes: According to the animal type to which the blood sample belongs, select a predetermined detection mode to detect the blood sample to obtain the first white blood cell histogram and the second white blood cell histogram.

23. A method for white blood cell classification and counting, characterized in that The method includes: Obtain a first white blood cell histogram of white blood cells in a blood sample treated with a hemolytic agent for a first treatment time and a second white blood cell histogram of white blood cells in a blood sample treated with the hemolytic agent for a second treatment time, wherein the second treatment time is greater than the first treatment time, the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value; Determine the peak shape of the second white blood cell histogram; When the peak shape of the second white blood cell histogram is bimodal, perform white blood cell classification and counting using the second white blood cell histogram; When the peak shape of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification results and counting results of the second white blood cell histogram and the classification results of the first white blood cell histogram.

24. A computer-readable storage medium, characterized in that, Comprises a program that can be executed by a processor to implement the method according to any one of claims 1 to 23.

25. A hematology analyzer, characterized in that, Comprises: A reaction cell; A sampling needle assembly for discharging a blood sample to be analyzed into the reaction cell; A hemolytic agent pushing member for pushing a hemolytic agent into the reaction cell, the hemolytic agent comprising a first hemolytic agent and a second hemolytic agent; A resistive detector for detecting white blood cells in a blood sample treated with the first hemolytic agent and generating a first signal, and for detecting white blood cells in a blood sample treated with the second hemolytic agent and generating a second signal; A processor for: Obtaining a first white blood cell histogram based on the first signal and obtaining a second white blood cell histogram based on the second signal, wherein the blood shadow value in the second white blood cell histogram is less than the blood shadow value in the first white blood cell histogram, and the blood shadow value in the second white blood cell histogram is less than a threshold blood shadow value; When the peak shape of the second white blood cell histogram is bimodal, perform white blood cell classification and counting using the second white blood cell histogram; When the peak shape of the second white blood cell histogram is unimodal, perform white blood cell classification and counting by combining the classification results and counting results of the second white blood cell histogram and the classification results of the first white blood cell histogram.

26. The hematology analyzer according to claim 25, wherein, The sampling needle assembly is used to discharge the blood sample to be analyzed into the reaction cell for the first time for the first measurement and to discharge the blood sample to be analyzed into the reaction cell for the second time for the second measurement; The hemolytic agent pushing member is used to: push the first hemolytic agent into the reaction cell to complete the first measurement and generate the first white blood cell histogram; Push the second hemolytic agent into the reaction cell to complete the second measurement and generate the second white blood cell histogram.

27. The blood analyzer according to claim 26, wherein The hemolytic agent pushing member is used to: First push the first hemolytic agent into the blood sample in the reaction cell to obtain a test solution, to complete the first measurement and obtain the first white blood cell histogram; Then push the second hemolytic agent into the test solution in the reaction cell to complete the second measurement and obtain the second white blood cell histogram.

28. The blood analyzer according to claim 25, wherein The hemolytic agent pushing component is used for: sequentially pushing at least three times of hemolytic agent into the same blood sample in the reaction cell, and each addition of hemolytic agent includes at least one of the first hemolytic agent and the second hemolytic agent; The resistive detector is used for: detecting white blood cells in the blood sample after each push of the first hemolytic agent and / or the second hemolytic agent by the hemolytic agent pushing component to generate corresponding signals; The processor is used for: generating a corresponding white blood cell histogram based on the corresponding signals, and selecting two successively generated white blood cell histograms in the corresponding white blood cell histogram as the first white blood cell histogram and the second white blood cell histogram respectively.

29. The hematology analyzer according to any one of claims 25-28, characterized in that, Performing white blood cell classification and counting by combining the classification and counting results of the second white blood cell histogram and the classification results of the first white blood cell histogram includes: Classifying and counting the second white blood cell histogram determined to be unimodal to obtain the counting result of white blood cells and the classification result of eosinophils and / or basophils in white blood cells; Obtaining a classification result of at least four classifications according to the classification result of the second white blood cell histogram and the classification result of the first white blood cell histogram that has been classified at least three times; Calculating the counting result of each type of white blood cell in the blood sample based on the classification result of at least four classifications and the counting result of white blood cells.

30. The hematology analyzer according to claim 29, wherein, The first white blood cell histogram that has been classified at least three times includes the first type of white blood cells, the second type of white blood cells, and the third type of white blood cells. Calculating the counting result of each type of white blood cell in the blood sample based on the classification result of at least four classifications and the counting result of white blood cells includes: Taking the counting result of eosinophils and / or basophils in the second white blood cell histogram as the counting result of eosinophils and / or basophils in the blood sample; Taking the counting result of the third type of white blood cells in the first white blood cell histogram that has been classified at least three times minus the counting result of eosinophils and / or basophils in the second white blood cell histogram as the counting result of the third type of white blood cells in the blood sample; Taking the counting result of the first type of white blood cells and the counting result of the second type of white blood cells in the first white blood cell histogram that has been classified at least three times as the counting result of the first type of white blood cells and the second type of white blood cells in the blood sample.

31. The blood analyzer according to claim 29, wherein The classification result of each type of white blood cell in the first white blood cell histogram is the proportion of each type of white blood cell in the first white blood cell histogram after removing the blood shadow area; and / or The classification result of eosinophils is the proportion of the area where eosinophils are located in the second white blood cell histogram; The classification result of basophils is the proportion of eosinophils in the second white blood cell histogram.

32. The hematology analyzer according to claim 25, wherein, It further includes a detection mode selection module for selecting a predetermined detection mode according to the animal type to which the blood sample belongs; the hemolytic agent pushing component pushes the corresponding hemolytic agent into the reaction cell for reaction according to the selected detection mode.

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