A method for detecting leukocytes in a blood cell analyzer

Abstract

This application discloses a white blood cell detection method for a blood cell analyzer. The method involves using a first detection module to detect white blood cell samples and obtain a white blood cell histogram. The count of basophils is then calculated from the white blood cell histogram. A second detection module then detects the white blood cell samples and obtains a scatter plot showing the two-dimensional distribution of four cell types, yielding the numbers of monocytes, neutrophils, and eosinophils. Finally, the five-part differential ratio of white blood cells is calculated based on the numbers of lymphocytes, monocytes, neutrophils, eosinophils, and basophils. This method can accurately detect various cell types within white blood cells. Furthermore, the calculation method is simple and cost-effective, thus achieving accurate detection results at a low cost.

Description

Technical Field

[0001] This invention belongs to the field of white blood cell detection technology, and particularly relates to a white blood cell detection method for a blood cell analyzer. Background Technology

[0002] Hematology analyzers are among the most widely used instruments in cell biology testing, with significant applications and research value in clinical practice, scientific research, and teaching. Some hematology analyzers can classify and count white blood cells, red blood cells, and platelets. Specifically, white blood cells include five types: lymphocytes, monocytes, neutrophils, eosinophils, and basophils. Therefore, they can be further classified and counted. White blood cell detection (such as white blood cell count and white blood cell differential) is one of the core functions of hematology analyzers and has always been a hot topic and challenge in engineering research.

[0003] Generally, high-end hematology analyzers can detect white blood cells more accurately. However, due to cost limitations, low-end hematology analyzers have lower accuracy and performance in detecting white blood cells compared to high-end analyzers. Taking white blood cell differential as an example, low-end hematology analyzers can generally only perform three-part differential analysis of white blood cells: small white blood cells (mainly lymphocytes), intermediate white blood cells (mainly monocytes, eosinophils, and basophils), and large white blood cells (mainly neutrophils). The monocytes, eosinophils, and basophils contained in the intermediate white blood cell group cannot be further effectively distinguished by low-end hematology analyzers. In contrast, high-end hematology analyzers can perform more accurate four- or five-part differential analysis of white blood cells.

[0004] As mentioned above, high-end hematology analyzers can detect white blood cells relatively accurately, but due to the high cost of both the equipment and the testing, they are not affordable for all hospitals and patients. On the other hand, low-end hematology analyzers are more popular with hospitals and patients because of their lower cost. However, as mentioned above, the performance and accuracy of low-end hematology analyzers in detecting white blood cells are somewhat compromised compared to high-end models, and they cannot meet the needs of high-end users or situations requiring precise results.

[0005] In some solutions, hospitals can purchase both high-end and low-end blood cell analyzers to solve this problem, but this is clearly not a better solution. Summary of the Invention

[0006] (I) Purpose of the Invention

[0007] To overcome the above shortcomings, the present invention aims to provide a white blood cell detection method for a blood cell analyzer, so as to solve the technical problem that the performance and accuracy of existing low-end blood cell analyzers in detecting white blood cells cannot meet the needs of high-end users or situations where the results require high precision.

[0008] (II) Technical Solution

[0009] To achieve the above objectives, the technical solution provided in this application is as follows:

[0010] A method for detecting white blood cells using a blood cell analyzer, comprising:

[0011] The white blood cell sample is detected based on the first detection module, and a white blood cell histogram is generated. The horizontal axis of the white blood cell histogram is the volume of each particle in the white blood cell, and the vertical axis is the number of particles in the corresponding volume. The white blood cell histogram sets the interference and small cell debris threshold line BA1 at the horizontal axis of 30, the basophil threshold line BA2 at the horizontal axis of 300, and the interference and large cell debris threshold line BA3 at the horizontal axis corresponding to the last data in the white blood cell histogram. The basophil count value is obtained by dividing the area between the basophil threshold line BA2 and the interference and large cell debris threshold line BA3 by the area between the interference and small cell debris threshold line BA1 and the basophil threshold line BA2.

[0012] Based on the second detection module, the white blood cell sample was detected to generate a two-dimensional scatter plot of four types of particle clusters, namely monocyte particle clusters, neutrophil particle clusters and eosinophil particle clusters.

[0013] This application uses a first detection module to detect white blood cell samples and obtain a white blood cell histogram. By calculating the white blood cell histogram, the count value of basophils is obtained. Then, a second detection module detects white blood cell samples and obtains a scatter plot of the two-dimensional distribution of four types of particle clusters, obtaining the number of monocytes, neutrophils, and eosinophils, respectively. Finally, the number of lymphocytes, monocytes, neutrophils, eosinophils, and basophils is obtained to calculate the five-part differential ratio of white blood cells. The above method can accurately detect various cell types in white blood cells. Moreover, the calculation method of this application is simple and has low computational cost, so accurate detection results can be obtained at a low cost.

[0014] In some embodiments, before detecting white blood cells based on the second detection module to obtain a scatter plot of the two-dimensional distribution of the four types of particle clusters, the method further includes:

[0015] The whole blood sample was thoroughly mixed with the staining agent and incubated at 35°C.

[0016] The stained sample is guided into the sheath flow cell;

[0017] Cell volume was determined by electrical resistance analysis of the sample in the sheath flow cell through a 60mm ruby ​​aperture.

[0018] The sample that has completed the resistance method analysis is introduced into the second sheath flow detection channel with a diameter of 42 mm for light absorption rate measurement to obtain information such as cell morphology and cell contents;

[0019] A two-dimensional scatter plot is generated based on cell size, cell morphology, and cell contents.

[0020] In some embodiments, the first detection module is provided with micro-perforations. Detecting white blood cells based on the first detection module and generating a white blood cell histogram includes: acquiring the magnitude of the corresponding electrical pulse value collected when each particle in the white blood cell sample passes through the micro-perforation and the frequency of each electrical pulse value; determining the volume of each particle according to the magnitude of the pulse value; calculating the number of particles based on the frequency of each pulse value; and generating a white blood cell histogram based on the volume of each particle and the number of particles. Attached Figure Description

[0021] Figure 1 This is a white blood cell histogram obtained by the white blood cell detection method of the blood cell analyzer of the present invention;

[0022] Figure 2 This is a scatter plot of the two-dimensional distribution obtained from the white blood cell detection method of the blood cell analyzer of the present invention. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments and the accompanying drawings. It should be understood that these descriptions are merely exemplary and not intended to limit the scope of the invention. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concept of the invention.

[0024] This invention provides a method for detecting white blood cells in a blood cell analyzer, comprising:

[0025] The white blood cell sample is detected based on the first detection module, and a white blood cell histogram is generated. The horizontal axis of the white blood cell histogram is the volume of each particle in the white blood cell, and the vertical axis is the number of particles in the corresponding volume. The white blood cell histogram sets the interference and small cell debris threshold line BA1 at the horizontal axis of 30, the basophil threshold line BA2 at the horizontal axis of 300, and the interference and large cell debris threshold line BA3 at the horizontal axis corresponding to the last data in the white blood cell histogram. The basophil count value is obtained by dividing the area between the basophil threshold line BA2 and the interference and large cell debris threshold line BA3 by the area between the interference and small cell debris threshold line BA1 and the basophil threshold line BA2.

[0026] The second detection module detects white blood cell samples to obtain a two-dimensional scatter plot of four types of particle clusters, and calculates the number of cells corresponding to the four types of particle clusters, which are monocyte particle clusters, neutrophil particle clusters and eosinophil particle clusters.

[0027] The five-part differential ratio of white blood cells is calculated based on the number of lymphocytes, monocytes, neutrophils, eosinophils, and basophils.

[0028] Specifically, the first detection module, also known as the BASO channel, is equipped with micro-perforations. The detection of white blood cells and the generation of a white blood cell histogram based on the first detection module includes: obtaining the magnitude of the corresponding electrical pulse value collected when each particle in the white blood cell sample passes through the micro-perforation and the frequency of each electrical pulse value; determining the volume of each particle according to the magnitude of the pulse value; calculating the number of each particle according to the frequency of each pulse value; and generating a white blood cell histogram based on the volume and number of each particle.

[0029] Specifically, the second detection module, also known as the DIFF channel, before detecting white blood cells based on the second detection module to obtain a scatter plot of the two-dimensional distribution of four types of particle clusters, also includes:

[0030] The whole blood sample was thoroughly mixed with the staining agent and incubated at 35°C.

[0031] The stained sample is guided into the sheath flow cell;

[0032] Cell volume was determined by electrical resistance analysis of the sample in the sheath flow cell through a 60mm ruby ​​aperture.

[0033] The sample that has completed the resistance method analysis is introduced into the second sheath flow detection channel with a diameter of 42 mm for light absorption rate measurement to obtain information such as cell morphology and cell contents;

[0034] A two-dimensional scatter plot is generated based on cell size, cell morphology, and cell contents.

[0035] Specifically, the purpose of thoroughly mixing the whole blood sample with the staining agent and incubating it at 35°C is:

[0036] ①Dissolve red blood cells;

[0037] ② The primary granules of monocytes, the specific granules of eosinophils and neutrophils were stained to different degrees, and the cell membranes (cell membrane, nuclear membrane and granule membrane) were also stained to different degrees.

[0038] ③ Fix the cell morphology to maintain its natural state. Because lymphocytes, monocytes, neutrophils and eosinophils stain differently with the dye, and the specific nuclear morphology and granule structure of each cell cause different light scattering intensities, a specific absorbance is produced.

[0039] Specifically, the stained samples are guided into the sheath flow chamber for dual sheath flow analysis. First, they pass through a 60mm ruby ​​aperture for resistivity analysis to determine cell size. Then, the samples quickly enter a second sheath flow detection channel with a diameter of 42mm for light absorption measurement to determine cell morphology and contents. Based on the characteristics of each cell type in these two analytical parameters, a two-dimensional scatter plot is generated. Within this channel, the classification of white blood cells into lymphocytes, monocytes, neutrophils, and eosinophils can be completed.

[0040] Preferably, this channel can detect two abnormal white blood cell subsets simultaneously with the intrinsic analysis: the percentage and absolute value of giant immature cells (LIC) and the percentage and absolute value of atypical lymphocytes (ALY).

[0041] Preferably, the first detection module uses a constant negative pressure impedance method to simulate cell size according to the size of each pulse in the electrical pulse signal. After the core algorithm, it can also obtain a histogram of the total number and volume distribution of white blood cells after removing interference.

[0042] Specifically, the blood cell analyzer equipped with the detection method of the present invention may further include: an interactive module, which is electrically connected to the first detection module and the second detection module. The user can select to start only the first detection module or start both the first detection module and the second detection module simultaneously through the interactive module, thus enabling different detection modes.

[0043] Low-precision mode: Only the first detection module is enabled to measure the number of white blood cells in the sample. The basophils in the sample are measured by impedance method. The first detection module is equipped with micro-perforations. Lymphocytes, monocytes, neutrophils, eosinophils and basophils in the sample pass through the micro-perforations to obtain the corresponding electrical pulse signals (first measurement data). The number of white blood cells is calculated according to the size of each pulse in the electrical pulse signal (particle volume) and its frequency of occurrence (number of particles corresponding to particle volume).

[0044] High-precision mode: Simultaneously activates the first and second detection modules, using the dual-detection module data verification algorithm; generates a histogram of total white blood cell count, white blood cell volume distribution, and a white blood cell LMNE scatter plot. Low-precision mode: Only activates the first detection module, generating a histogram of total white blood cell count and white blood cell volume distribution.

[0045] It should be understood that the specific embodiments described above are merely illustrative or explanatory of the principles of the invention and do not constitute a limitation thereof. Therefore, any modifications, equivalent substitutions, improvements, etc., made without departing from the spirit and scope of the invention should be included within the protection scope of the invention. Furthermore, the appended claims are intended to cover all variations and modifications falling within the scope and boundaries of the appended claims, or equivalent forms of such scope and boundaries.

Claims

1. A method of detecting leukocytes in a blood cell analyzer, characterized by, The method comprises the following steps: detecting the white blood cell sample based on the first detection module and generating a white blood cell histogram, wherein the abscissa of the white blood cell histogram is the volume of each particle in the white blood cell, the ordinate is the number of particles corresponding to the volume, and in the white blood cell histogram: a disturbance and small cell fragment threshold line BA1 is set at the position of the abscissa 30; an alkaline granulocyte threshold line BA2 is set at the position of the abscissa 300; a disturbance and large cell fragment threshold line BA3 is set at the position of the abscissa corresponding to the end of the white blood cell histogram data; the area value formed between the alkaline granulocyte threshold line BA2 and the disturbance and large cell fragment threshold line BA3 is divided by the area value between the disturbance and small cell fragment threshold line BA1 and the alkaline granulocyte threshold line BA2 to directly obtain the count value of the alkaline granulocyte; detecting the white blood cell sample based on the second detection module, generating a two-dimensional distribution scatter plot of four types of particle groups, and calculating the number of cells corresponding to the four types of particle groups respectively, wherein the four types of particle groups include lymphocyte particle groups, monocyte particle groups, neutrophil particle groups and eosinophil particle groups; based on the count value of the alkaline granulocyte and the number of cells corresponding to the four types of particle groups respectively, calculating the white blood cell five-classification ratio.

2. The method of leucocyte detection of a blood cell analyzer according to claim 1, wherein Before the detection of the white blood cell sample based on the second detection module, the method further comprises the following steps: fully mix and incubate the whole blood sample with the staining agent at 35 DEG C; introduce the sample after staining into a sheath flow cell; make the sample pass through a 60mm ruby orifice, and obtain the cell volume by resistance method analysis; introduce the sample after resistance method analysis into a second sheath flow detection channel with a diameter of 42mm, and obtain the cell morphology and cell content information by light absorption rate determination; generate the two-dimensional distribution scatter plot according to the cell volume, the cell morphology and the cell content information.

3. The method of claim 1, wherein the step of detecting the white blood cells comprises the steps of: detecting the white blood cells in the blood sample by using a light source; and detecting the white blood cells in the blood sample by using a light source. The first detection module is provided with a microvia, and the detection of the white blood cell sample based on the first detection module and the generation of the white blood cell histogram comprise the following steps: obtain the size of the electric pulse value generated by each particle in the white blood cell sample when passing through the microvia and the frequency of each electric pulse value; determine the volume of each particle according to the size of the pulse value; calculate the number of particles corresponding to the volume according to the frequency of each electric pulse value; generate the white blood cell histogram according to the volume of each particle and the corresponding number of particles.

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