CN110887818A

CN110887818A - Blood sample analysis method, blood cell analyzer and storage medium

Info

Publication number: CN110887818A
Application number: CN201811045480.8A
Authority: CN
Inventors: 祁欢; 王官振
Original assignee: Shenzhen Mairui Technology Co Ltd; Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mairui Technology Co Ltd; Shenzhen Mindray Bio Medical Electronics Co Ltd
Priority date: 2018-09-07
Filing date: 2018-09-07
Publication date: 2020-03-17
Anticipated expiration: 2038-09-07
Also published as: CN110887818B

Abstract

A blood sample analysis method, a blood cell analyzer and a storage medium, which are used for acquiring fluorescence signal information of a blood sample treated by a fluorescent reagent; obtaining a measured parameter of red blood cells in the blood sample; and correcting the fluorescence signal information according to the erythrocyte parameters and/or sending out a fluorescence signal information abnormity prompt.

Description

Blood sample analysis method, blood cell analyzer and storage medium

Technical Field

The invention relates to the field of sample analysis, in particular to an analysis method of a blood sample, a blood cell analyzer and a storage medium.

Background

A blood cell analyzer is an instrument that can detect cells in blood, and can classify and count white blood cells, red blood cells, platelets, nucleated red blood cells, reticulocytes, and the like in blood.

Taking leukocyte detection as an example, the most common method is laser scattering, i.e., a blood cell analyzer uses scattered light to classify and count leukocytes. The scattered light comprises three optical signals of forward scattered light, side scattered light, fluorescent signal and the like. Generally, the forward scattered light can reflect the size information of the cell, the side scattered light can reflect the complexity of the internal structure of the cell, and the fluorescent signal can reflect the content of DNA, RNA, etc. in the cell, which can be stained by the fluorescent dye. The white blood cells can be classified by using the optical signals, and the count value of the white blood cells can be obtained.

When the existing leukocyte differential counting method is applied to some blood detection, the accuracy cannot meet the requirement. Therefore, there is a need to develop a method for classifying leukocytes with better accuracy.

Disclosure of Invention

Provided are a method of analyzing a blood sample, a blood cell analyzer, and a storage medium.

There is provided a method of analyzing a blood sample, comprising:

obtaining fluorescence signal information of the blood sample treated by the fluorescent reagent;

obtaining a measured parameter of red blood cells in the blood sample;

and correcting the fluorescence signal information according to the erythrocyte parameters and/or sending out a fluorescence signal information abnormity prompt.

In one embodiment, the acquiring fluorescence signal information of the blood sample processed by the fluorescence reagent includes: and acquiring fluorescence signal information, forward scattered light signal information and/or side scattered light signal information of the blood sample treated by the fluorescent reagent.

In one embodiment, the acquiring fluorescence signal information of the blood sample processed by the fluorescence reagent includes: and measuring the blood sample by adopting flow cytometry to acquire fluorescence signal information of the blood sample treated by the fluorescent reagent, and forward scattering light signal information and/or side scattering light signal information.

In one embodiment, the method further comprises the following steps: and carrying out white blood cell classification and/or counting according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the method further comprises the following steps: and counting the nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the method further comprises the following steps: and performing white blood cell counting and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the method further comprises the following steps: and performing basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, before acquiring the fluorescence signal information of the blood sample processed by the fluorescence reagent, the method further includes: the method includes performing a red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, and then performing a hemolysis process and a fluorescent reagent process on the blood sample and measuring fluorescence signal information of the blood sample.

In one embodiment, the performing a red blood cell parameter measurement on the blood sample to determine the red blood cell parameter comprises: measuring a red blood cell parameter of the blood sample using a pinhole impedance method or an optical method to determine the red blood cell parameter.

In one embodiment, the correcting the fluorescence signal information according to the red blood cell parameter includes: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as dependent variables.

In one embodiment, the correction function is an increasing function of the dependent variable relative to the independent variable.

In one embodiment, the modification function is a linear function, a polynomial function, or a power function.

In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.

In one embodiment, after the correcting the fluorescence signal information according to the red blood cell parameter, the method further includes: and outputting the corrected fluorescence signal information.

In one embodiment, after the correcting the fluorescence signal information according to the red blood cell parameter, the method further includes: and outputting the fluorescence signal information before and after correction.

In one embodiment, after the correcting the fluorescence signal information according to the red blood cell parameter, the method further includes: and sending out a prompt that the fluorescence signal information is corrected or sending out an alarm.

In one embodiment, the indication that the fluorescence signal information has been modified comprises: and outputting the corrected amplitude of the fluorescence signal information.

In one embodiment, after the correcting the fluorescence signal information according to the red blood cell parameter, the method further includes: and if the correction amplitude of the fluorescence signal information exceeds a set threshold value, sending out a prompt that the fluorescence signal information is corrected or sending out an alarm.

There is also provided a blood cell analyzer including:

at least one reaction cell for providing a reaction site for the blood sample and the reagent;

the optical detection device is used for irradiating the blood sample treated by the reagent by light, collecting optical signals generated by each particle in the blood sample treated by the reagent due to the light irradiation, and converting the optical signals into electric signals so as to output optical signal information;

the conveying device is used for conveying the blood sample treated by the reagent in the reaction pool to the optical detection device;

the processor is used for receiving and processing the optical signal information output by the optical detection device to obtain a measurement parameter of the blood sample; the processor obtains fluorescence signal information of the blood sample processed by the fluorescence reagent, obtains measured erythrocyte parameters in the blood sample, and corrects the fluorescence signal information and/or sends out a fluorescence signal information abnormal prompt according to the erythrocyte parameters.

In one embodiment, the processor obtains fluorescence signal information of the blood sample treated with the fluorescent reagent, and forward scattered light signal information and/or side scattered light signal information of leukocyte particles.

In one embodiment, the optical detection device includes a light source and a flow chamber having an aperture, leukocyte particles in the blood sample treated by the fluorescent reagent flow in the flow chamber and pass through the aperture one by one, light emitted by the light source irradiates the particles in the aperture and generates a fluorescent signal and a scattered light signal correspondingly, and the fluorescent signal information, the forward scattered light signal information and/or the side scattered light signal information are output to the processor after conversion by the optical detection device.

In one embodiment, the processor further performs leukocyte classification and/or enumeration based on the side scattered light values and the corrected fluorescence signal information of the blood sample.

In one embodiment, the processor further performs a nucleated red blood cell count based on the forward scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the processor further performs white blood cell counting and/or reticulocyte recognition and/or basophil classification based on the forward scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the processor further performs basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

In one embodiment, the reaction cell provides a reaction site for the blood sample and a reagent required for measuring the erythrocyte parameter, the optical detection device measures the erythrocyte parameter of the blood sample treated by the reagent and outputs optical signal information, and the processor determines the erythrocyte parameter according to the optical signal information; the reaction cell carries out hemolysis treatment and fluorescence reagent treatment on the blood sample, then the optical detection device measures the treated blood sample and outputs the fluorescence signal information of the blood sample.

In one embodiment, the processor is configured to: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as dependent variables.

In one embodiment, the processor further outputs the corrected fluorescence signal information.

A computer-readable storage medium storing a program executable by a processor to implement a method as in any one of the embodiments above.

According to the blood sample analysis method, the blood cell analyzer and the computer-readable storage medium of the above embodiments, the fluorescence signal information is corrected using the red blood cell parameter, and the accuracy of the fluorescence signal information is improved.

Drawings

FIG. 1 is a schematic view showing the structure of a blood cell analyzer according to an embodiment;

FIG. 2 is a flow chart of a method of analyzing a blood sample according to one embodiment;

FIG. 3 is a flow chart of a method of analyzing a blood sample according to another embodiment;

FIG. 4 is a flow chart of a method of analyzing a blood sample according to yet another embodiment;

FIG. 5 is a schematic structural diagram of a fluorescence signal information correction apparatus for a blood sample according to an embodiment;

FIG. 6 is a scatter plot of the position values of WBC clusters in the fluorescence direction versus HCT for a plurality of samples;

FIG. 7 is a scatter plot of the location values of WBC clusters in the fluorescence direction versus RBC statistics for multiple samples;

FIG. 8 is a scattergram of statistical location values of WBC clusters in the fluorescence direction versus HGB in multiple samples;

FIG. 9 is a functional diagram of a first function modified using HCT according to one embodiment;

FIG. 10 is a graph showing fluorescence signal information of cell particles before and after correction in three samples.

Detailed Description

The present invention will be described in further detail with reference to the following detailed description and accompanying drawings. Wherein like elements in different embodiments are numbered with like associated elements. In the following description, numerous details are set forth in order to provide a better understanding of the present application. However, those skilled in the art will readily recognize that some of the features may be omitted or replaced with other elements, materials, methods in different instances. In some instances, certain operations related to the present application have not been shown or described in detail in order to avoid obscuring the core of the present application from excessive description, and it is not necessary for those skilled in the art to describe these operations in detail, so that they may be fully understood from the description in the specification and the general knowledge in the art.

Furthermore, the features, operations, or characteristics described in the specification may be combined in any suitable manner to form various embodiments. Also, the various steps or actions in the method descriptions may be transposed or transposed in order, as will be apparent to one of ordinary skill in the art. Thus, the various sequences in the specification and drawings are for the purpose of describing certain embodiments only and are not intended to imply a required sequence unless otherwise indicated where such sequence must be followed.

The numbering of the components as such, e.g., "first", "second", etc., is used herein only to distinguish the objects as described, and does not have any sequential or technical meaning. The term "connected" and "coupled" when used in this application, unless otherwise indicated, includes both direct and indirect connections (couplings).

Taking a common five-classification blood cell analyzer as an example, the device generally has a channel for measuring a fluorescence signal, and the device uses three scattered light signals, such as forward scattered light, side scattered light, and a fluorescence signal, to classify and count cells, such as white blood cell count, nucleated red blood cell count, and some basophil cells.

Some english terms of the present application will be explained first.

RBC: red blood cells (number);

NRBC: nucleated red blood cells (number);

RET: reticulocytes (number);

HGB: red blood cell hemoglobin content;

HCT: hematocrit;

MCV: mean red blood cell volume;

WBC: white blood cells (number);

BASO: basophils (number).

The conventional five-classification blood cell analyzer has a channel for measuring fluorescence signals, and the channel uses three optical signals, such as forward scattered light, lateral fluorescence signals and the like, to classify and count cells, such as white blood cell count, nucleated red blood cell classification and basophil classification. The channel herein refers to a detection channel for identifying and/or counting and/or classifying a certain type of cells, for example, the NRBC channel is a channel for counting nucleated red blood cells, the WNB channel is a channel for classifying nucleated red blood cells, the RET channel is a channel for classifying and/or counting reticulocytes, the DIFF channel is a channel for classifying and/or counting white blood cells, and the BASO channel is a channel for counting basophils, which are known to those skilled in the art and will not be described herein.

By obtaining the optical signals of the cells for classification and counting, classification errors occur when WBC clusters of different samples appear at different positions. In leukocyte classification, the applicant has found through long-term development that in some samples, when cells are treated with a fluorescent dye and classified using a fluorescent signal and a scattered light signal, a case occurs in which the measurement result deviates from the result of microscopic examination. It has been found through intensive studies that the parameters of erythrocytes in such samples differ from normal values. Through verification, the red blood cell parameters and the fluorescence signal intensity have certain correlation, and the fluorescence signals can be corrected by using the red blood cell parameters. While not being bound by theory, applicants speculate that red blood cells may affect the staining behavior of other cells that need to be sorted, resulting in an abnormal fluorescence signal.

In practice, the applicant finds that, in the currently common five-classification blood cell analyzer, for example, the NRBC channel is not taken as an example, when a cell is stained with a fluorescent dye, positions of fluorescence signals of cells in a fluorescence direction of different samples may be different, and in a process of counting and classifying cells (e.g., white blood cells), an excessive or an insufficient fluorescence signal may affect a counting and classifying gating white blood cell algorithm (e.g., the fluorescence signal affects gating), which may affect counting and classifying of the NRBC channel, and further may increase a risk of clinical misdiagnosis. For the RET channel, the cells are treated with different fluorescent dyes due to different distribution of red blood cells, so that the signals of the cells in the fluorescence direction are different, thereby affecting the classification of red blood cells, such as the identification of RET.

Therefore, for those channels using fluorescent dyes, such as NRBC channel, RET channel, and DIFF channel, when these channels are used for sample measurement, if the fluorescent signal correction is not performed, the fluorescent signal of WBC particles will be too large or too small, and in the case of limited byte number of the signal collector, because the signal will have maximum and minimum limits, the WBC particle clusters will be too left or right in the scatter diagram, thereby resulting in distortion of the channel scatter diagram, missing necessary scatter diagram information, causing difficulty in subsequent classification, and easily causing erroneous classification of blood shadow, nucleated red blood cells, and basophils, thereby giving erroneous counting and classification results.

In summary, for those channels that use fluorescent dyes, such as NRBC channel, BASO channel, WNB channel, RET channel, and DIFF channel, a solution for correcting the difference in fluorescence signals of different samples in these channels is needed.

In order to solve the above problems, the applicant has invented the research of a large number of samples through reagent theory of action analysis, and in the actual measurement process of a channel using a fluorescent dye, such as an NRBC channel, the fluorescence signal of WBC particles is in a negative correlation with red blood cell count (RBC), red blood cell hemoglobin content (HGB), and red cell Hematocrit (HCT), that is, the smaller the RBC, HGB, and HCT is, the larger the fluorescence signal of WBC particles is; the larger the RBC, HGB, HCT, the smaller the WBC particle fluorescence signal. Therefore, the RBC, the HGB and the HCT can be used for correcting the fluorescence signals of the WBC particles and/or sending out the fluorescence signal information abnormity prompt, namely, the fluorescence signal information can be corrected and/or the fluorescence signal information abnormity prompt can be sent out according to the red blood cell parameters. Only the correction may be performed, only the abnormality indication may be issued, and both the correction and the abnormality indication may be issued.

Example 1

The blood cell analyzer according to an embodiment of the present invention may mainly include a structure as shown in fig. 1: at least one reaction cell 101, an optical detection device 102, a transport device 103, and a processor 104, as described in detail below.

The reaction cell 101 is used for providing a reaction site for the blood sample and the reagent to prepare a sample solution. Specifically, a blood sample obtained by blood collection may be diluted and labeled with a fluorescent staining reagent to obtain a sample solution. Commonly used fluorescent staining reagents may be pyronine, acridine orange, thiazole orange, and the like.

The optical detection device 102 is configured to irradiate the blood sample treated by the reagent, that is, the sample liquid, with light, collect optical signals generated by light irradiation of each particle in the blood sample treated by the reagent, convert the optical signals into electrical signals, and output optical signal information. The optical signal may be a forward scattered light signal, a side scattered light signal, or a fluorescent signal. In one embodiment of the optical detection apparatus 102, which may include a light source 1021 and a sheath flow chamber 1022 having an aperture 10221, etc., particles in a blood sample may flow within the sheath flow chamber 1022 and pass through the aperture 10221 one by one, and light emitted from the light source 1021 may illuminate the particles in the aperture 10221 and generate a scattered light signal and/or a fluorescent light signal, respectively. The optical detection device 102 may further include a lens set 1023, a photo sensor 1024 (such as a photodiode, a photomultiplier tube, etc.) and an a/D converter respectively disposed in front of and laterally to the aperture, the a/D converter may be disposed in the processor 104 or separately form one element, so that the lens set 1023 may capture the corresponding scattered light signal and the fluorescence signal, the photo sensor 1024 may convert the captured optical signal (such as the scattered light signal and the fluorescence signal) into an electrical signal, and the a/D converter may perform a/D conversion on the electrical signal to obtain a digital signal, and the digital signal may be output as optical signal information.

The transport device 103 is used to transport the blood sample treated with the reagent in the reaction cell 101, i.e., the sample liquid, to the optical detection device 102.

The processor 104 is used for receiving and processing the optical signal information output by the optical detection device 102 to obtain the measured parameter of the blood sample. In one embodiment, the processor 104 obtains fluorescence signal information of a blood sample processed by a fluorescence reagent, obtains a measured red blood cell parameter of the blood sample, and corrects the fluorescence signal information according to the red blood cell parameter. The particles represented by the fluorescence signal are mainly leukocyte particles or ghost particles, wherein the ghost particles are particles with the original shape and size of the cell membrane structure remained after the rupture of the plasma membrane after the hypotonic treatment of the red blood cells. In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.

As mentioned above, the data obtained by the processor 104 includes information on the fluorescence signal of the blood sample and the parameters of the red blood cells in the blood sample, and in one embodiment, the processor 104 can also obtain information on the forward scattered light signal or the side scattered light signal of the blood sample. These three types of data-first type of data: fluorescence information number information of blood sample, second type data: forward scattered light signal information or side scattered light signal information of the blood sample, and a third type of data: one or more of the parameters of red blood cells in the blood sample may be transmitted by other devices to the processor of the blood cell analyzer of the present invention, or may be measured by the blood cell analyzer of the present invention, as exemplified below.

Acquisition of the first type of data (fluorescence information number information of the blood sample) by the processor 104: the reaction cell 101 performs hemolysis treatment and fluorescence reagent treatment on the blood sample, the transportation device 103 transports the hemolyzed blood sample and the fluorescence reagent treated blood sample to the optical detection device 102, the optical detection device 102 measures the hemolyzed blood sample and the fluorescence reagent treated blood sample, and outputs fluorescence signal information of the blood sample to the processor 104.

Acquisition of a second type of data (forward scatter signal information or side scatter signal information of the blood sample) by processor 104: forward scattered light signal information and side scattered light signal information can be obtained when the fluorescence signal is obtained; for example, the reaction cell 101 performs a hemolysis process and a fluorescence reagent process on a blood sample, the transportation device 103 transports the hemolyzed blood sample and the fluorescence reagent processed blood sample to the optical detection device 102, the optical detection device 102 measures the hemolyzed blood sample and the fluorescence reagent processed blood sample, and outputs fluorescence signal information, forward scattered light signal information and/or side scattered light signal information of the blood sample to the processor 104. Specifically, by the transportation of the transportation device 103, the cell particles in the blood sample processed by the fluorescent reagent flow in the sheath flow chamber 1022 and pass through the orifice 10221 one by one, the light emitted by the light source 1021 irradiates the cell particles in the orifice 10221 and generates the fluorescent signal and the scattered light signal (referring to one or both of the forward scattered light signal information and the side scattered light signal information) of the cell particles correspondingly, and outputs the fluorescent signal information and the forward scattered light signal information or the side scattered light signal information to the processor 104 after the conversion of the optical detection device 10221.

The processor 104 can perform the following cell detection by using the acquired fluorescence signal information and the acquired emission light signal information. The white blood cells are classified and/or counted according to the side scattered light value of the blood sample and the corrected fluorescence signal information, for example, the white blood cells are classified four by the DIFF channel. The nucleated red blood cell count is performed based on the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, by the NRBC channel. And (3) carrying out white blood cell counting and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, carrying out reticulocyte classification and/or counting through a RET channel. And performing basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information, for example, performing the basophil classification through a BASO channel.

Acquisition of a third type of data (red blood cell parameters in the blood sample) by processor 104: the reaction cell 101 provides a reaction site for the blood sample and a reagent required for measuring the erythrocyte parameter, the conveying device 103 conveys the blood sample processed by the reagent to the optical detection device 102, the optical detection device 102 measures the erythrocyte parameter of the blood sample processed by the reagent and outputs optical signal information, and the processor 104 determines the erythrocyte parameter according to the optical signal information. I.e. parameters of red blood cells in a sample of blood obtained can be detected optically. Of course, in other embodiments, the red blood cell parameter in the blood sample can also be obtained by a small-hole impedance measurement, for example, by measuring the blood sample by the small-hole impedance measurement from the detection device of the blood cell analyzer itself or other instruments.

The above is a description of the acquisition of the above three types of data by the processor 104, and the following is a description of the modification of the processor 104.

In one embodiment, the processor 104 corrects the fluorescence signal information according to the red blood cell parameter, which may include: and performing increase correction and/or decrease correction on the fluorescence signal information according to the erythrocyte parameters. The goal of the correction is: the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle no matter how the red blood cell parameters of HCT, RBC and HGB are changed.

(1) Augmentation correction

The processor 104 performs an augmentation correction on the fluorescence signal information according to the red blood cell parameter, including: and if the erythrocyte parameter is larger than a first preset value, performing augmentation correction on the fluorescence signal information. Further, in one embodiment, if the red blood cell parameter is greater than the first predetermined value, the greater the red blood cell parameter, the greater the magnitude of the incremental correction performed by the processor 104 on the fluorescence signal information. The first preset value can be set as a reference according to the normal red blood cell parameter value under the condition of normal fluorescence intensity of the particles.

As described above, the red blood cell parameters may include at least one of RBC, HGB, and HCT. When the red blood cell parameter includes only one of RBC, HGB, and HCT, then it is well understood that the red blood cell parameter is greater than the first preset value; when the red blood cell parameter includes two or three of RBC, HGB, and HCT, the red blood cell parameter being greater than the first preset value may refer to: and performing weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameters, wherein the specific weight value can be flexibly set in practice, the calculated value is used as the red blood cell parameter to be compared with a first preset value, and when the calculated value is larger than the first preset value, the red blood cell parameter is considered to be larger than the first preset value. For example, when the red blood cell parameters include three of RBC, HGB and HCT, the three parameters of RBC, HGB and HCT are calculated to perform weighted sum or weighted average calculation, the calculated value is regarded as the value of the red blood cell parameter to be compared with a first preset value, and when the calculated value is greater than the first preset value, the red blood cell parameter is greater than the first preset value.

(2) Reduction correction

Processor 104 performs a subtraction correction on the fluorescence signal information according to the red blood cell parameters, including: and if the red blood cell parameter is smaller than a first preset value, performing reduction correction on the fluorescence signal information. Further, in one embodiment, if the red blood cell parameter is less than the first predetermined value, the smaller the red blood cell parameter, the larger the magnitude of the decrease correction of the fluorescence signal information by the processor 104

As described above, the red blood cell parameters may include at least one of RBC, HGB, and HCT. When the red blood cell parameter includes only one of RBC, HGB, and HCT, then it is well understood that the red blood cell parameter is less than the first preset value; when the red blood cell parameter includes two or three of RBC, HGB, and HCT, the red blood cell parameter being less than the first preset value may refer to: and performing weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameters, wherein the specific weight value can be flexibly set in practice, the calculated value is used as the red blood cell parameter to be compared with a first preset value, and when the calculated value is smaller than the first preset value, the red blood cell parameter is considered to be smaller than the first preset value. For example, when the red blood cell parameters include three of RBC, HGB and HCT, the three parameters of RBC, HGB and HCT are calculated to perform weighted sum or weighted average calculation, the calculated value is regarded as the value of the red blood cell parameter to be compared with the first preset value, and when the calculated value is smaller than the first preset value, it indicates that the red blood cell parameter is smaller than the first preset value.

Processor 104 performs an increase correction and/or a decrease correction on the fluorescence signal information according to the red blood cell parameters, including: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as dependent variables, wherein the correction function is an increasing function of the dependent variables relative to the independent variables.

In one embodiment, the modification function may be a linear function or a power function. In one embodiment, the modification function includes a first function and a second function, and both the first function and the second function are increasing functions; the processor substituting the red blood cell parameters and the fluorescence signal information as independent variables into a preset correction function comprises: substituting the erythrocyte parameter as an independent variable into the first function to obtain a correction coefficient, and substituting the correction coefficient as a constant and the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information as a dependent variable. In one embodiment of the processor 104, the corrected fluorescence signal information may also be output, such as displaying a fluorescence signal scatter plot on a display based on the corrected fluorescence signal information, or sending the corrected fluorescence signal information to a remote server, a central station. Of course, in some embodiments, the fluorescence signal information before and after correction may also be output, for example, a fluorescence signal scatter diagram after correction is displayed on the display according to the fluorescence signal information after correction, and a fluorescence signal scatter diagram before correction is displayed on the display according to the fluorescence signal information before correction, so as to facilitate comparison by the user.

In some embodiments, after the correction, the processor 104 may also issue a prompt or alert that the fluorescent signal information has been corrected, such as by issuing a prompt to the user via a display. In some embodiments, the processor 104 may also output the corrected magnitude of the fluorescence signal information, for example, by displaying the corrected magnitude of the fluorescence signal information in percentage form via a display. In some embodiments, the indication that the fluorescence signal information has been modified or the alarm is issued is only issued if the magnitude of the modification of the fluorescence signal information exceeds a set threshold.

The correction function, the first function and the second function are exemplified below.

In the formulae of the following formulae (1) to (7), y_flRepresenting the corrected fluorescence signal information, x_flRepresenting fluorescence signal information before correction, x₁、x₂、x₃The values of HCT, RBC and HGB are shown, respectively.

(1) When the red blood cell parameters include only HCT, the correction function may have the form:

y_fl＝f₁(x₁,x_fl)；

further, a correction function f₁(x₁,x_fl) The first function included may be f₁₁(x₁) The second function may be f₁₂(x_fl) E.g. second function f₁₂(x_fl)＝k₁x_fl+b，k₁And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₁＝f₁₁(x₁)。

(2) When the red blood cell parameter includes only RBCs, the correction function may have the form:

y_fl＝f₂(x₂,x_fl)；

further, a correction function f₂(x₂,x_fl) The first function included may be f₂₁(x₂) The second function may be f₂₂(x_fl) E.g. second function f₂₂(x_fl)＝k₂x_fl+b，k₂And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₂＝f₂₁(x₂)。

(3) When the red blood cell parameters include only HGBs, the correction function may have the form:

y_fl＝f₃(x₃,x_fl)；

further, a correction function f₃(x₃,x_fl) The first function included may be f₃₁(x₃) The second function may be f₃₂(x_fl) E.g. second function f₃₂(x_fl)＝k₃x_fl+b，k₃And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₃＝f₃₁(x₃)。

(4) When the red blood cell parameters include HCT and RBC, the correction function can have the form:

y_fl＝f₄(x₁,x₂,x_fl)；

further, a correction function f₄(x₁,x₂,x_fl) The first function included may be f₄₁(x₁,x₂) The second function may be f₄₂(x_fl) E.g. second function f₄₂(x_fl)＝k₄x_fl+b，k₄And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₄＝f₄₁(x₁,x₂)。

(5) When the red blood cell parameters include RBC and HGB, the correction function may have the form:

y_fl＝f₅(x₂,x₃,x_fl)；

further, a correction function f₅(x₂,x₃,x_fl) The first function included may be f₅₁(x₂,x₃) The second function may be f₅₂(x_fl) E.g. second function f₅₂(x_fl)＝k₅x_fl+b，k₅And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₅＝f₅₁(x₂,x₃)。

(6) When the red blood cell parameters include HCT and HGB, the correction function may have the form:

y_fl＝f₆(x₁,x₃,x_fl)；

further, a correction function f₆(x₁,x₃,x_fl) The first function included may be f₆₁(x₁,x₃) The second function may be f₆₂(x_fl) E.g. second function f₆₂(x_fl)＝k₆x_fl+b，k₆And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₆＝f₆₁(x₁,x₃)。

(7) When the red blood cell parameters include HCT, RBC, and HGB, the correction function may have the form:

y_fl＝f₇(x₁,x₂,x₃,x_fl)；

further, a correction function f₇(x₁,x₂,x₃,x_fl) Comprises a firstThe function may be f₇₁(x₁,x₂,x₃) The second function may be f₇₂(x_fl) E.g. second function f₇₂(x_fl)＝k₇x_fl+b，k₇And b is a constant, and the substitution of the correction factor as a constant into the second function may mean k₇＝f₇₁(x₁,x₂,x₃)。

Example 2

An embodiment of the present invention further provides a method for analyzing a blood sample (hereinafter referred to as an analysis method). Referring to FIG. 2, the analysis method may include steps 310 to 350.

Step 310: and acquiring fluorescence signal information of the blood sample treated by the fluorescent reagent. In one embodiment, step 310 may comprise: and acquiring fluorescence signal information of the blood sample treated by the fluorescent reagent, and forward scattered light signal information or side scattered light signal information.

Step 330: obtaining a measured parameter of red blood cells in the blood sample. In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.

In the above steps, the data of the fluorescence signal information, the forward scattering light signal information and the side scattering light signal information obtained in step 310 may be measured in real time or may be measured in non-real time, for example, the data is stored in a memory, and step 310 obtains the data from the memory. Similarly, the red blood cell parameters obtained in step 330 may be obtained by real-time measurement or non-real-time measurement, for example, the red blood cell parameters in the blood sample are stored in a memory in advance, and step 330 obtains the red blood cell parameters in the blood sample from the memory.

Referring to fig. 3, instead of taking the real-time measurement of the fluorescence signal information of the blood sample and the parameters of the red blood cells in the blood sample as an example, an embodiment of the analysis method may further include

steps

300 and 320, which are described in detail below.

Step 300: subjecting the blood sample to hemolysis treatment and fluorescent reagent treatment, and measuring fluorescence signal information of the blood sample.

Step 320: performing a red blood cell parameter measurement on the blood sample to determine the red blood cell parameter. In one embodiment, if the analysis method has both

steps

300 and 320, step 320 may be performed first, and then step 300 may be performed, for example, performing a red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, then performing a hemolysis process and a fluorescence reagent process on the blood sample, and measuring the fluorescence signal information of the blood sample.

Instead of obtaining fluorescence signal information, scattered light signal information (forward scattered light signal information or side scattered light signal information) of the blood sample in real-time measurement, in one embodiment of the analysis method, step 310 may be measuring the blood sample using flow cytometry to obtain fluorescence signal information, and forward scattered light signal information or side scattered light signal information of the blood sample after being treated with the fluorescent reagent.

By the acquired fluorescence signal information and the acquired scattered light signal information, the following cell detection can be performed. The white blood cells are classified and/or counted according to the side scattered light value of the blood sample and the corrected fluorescence signal information, for example, the white blood cells are classified four by the DIFF channel. The nucleated red blood cell count is performed based on the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, by the NRBC channel. And (3) carrying out white blood cell counting and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, carrying out reticulocyte classification and/or counting through a RET channel. And performing basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information, for example, performing the basophil classification through a BASO channel.

Step 350: and correcting the fluorescence signal information according to the erythrocyte parameters and/or sending out a fluorescence signal information abnormity prompt. Namely, the fluorescence signal information can be corrected and/or the fluorescence signal information abnormity prompt can be sent out according to the erythrocyte parameters. Only the correction or the abnormal prompt may be issued, or both the correction and the abnormal prompt may be issued

In some embodiments, step 350 modifies the fluorescence signal information according to the red blood cell parameters, including: and performing increase correction and/or decrease correction on the fluorescence signal information according to the erythrocyte parameters. The goal of the correction is: the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle no matter how the red blood cell parameters of HCT, RBC and HGB are changed.

(1) Augmentation correction

Step 350, performing augmentation correction on the fluorescence signal information according to the red blood cell parameters, including: and if the erythrocyte parameter is larger than a first preset value, performing augmentation correction on the fluorescence signal information. Further, in an embodiment, if the red blood cell parameter is greater than the first preset value, the larger the red blood cell parameter is, the larger the amplitude of the increase correction performed on the fluorescence signal information by step 350 is. The first preset value can be set as a reference according to the normal red blood cell parameter value under the condition of normal fluorescence intensity of the particles.

(2) Reduction correction

Step 350, performing a reduction correction on the fluorescence signal information according to the red blood cell parameter, including: and if the red blood cell parameter is smaller than a first preset value, performing reduction correction on the fluorescence signal information. Further, in one embodiment, if the red blood cell parameter is smaller than the first predetermined value, the smaller the red blood cell parameter, the larger the amplitude of the reduction correction of the fluorescence signal information in step 350 is

Step 350, performing an increase correction and/or a decrease correction on the fluorescence signal information according to the red blood cell parameter, including: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information, wherein the correction function is an increasing function. In one embodiment, the modification function is a linear function or a power function. In one embodiment, the modification function includes a first function and a second function, and both the first function and the second function are increasing functions; the processor substituting the red blood cell parameters and the fluorescence signal information as independent variables into a preset correction function comprises: substituting the erythrocyte parameter as an independent variable into the first function to obtain a correction coefficient, and substituting the correction coefficient as a constant and the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information. In one embodiment, the first function and the second function are both linear functions. For examples of the modification function, the first function and the second function, reference may be made to embodiment 1, and details will be given in the following embodiments for examples, which are not described herein again.

Referring to fig. 4, an embodiment of the analysis method may further include step 370: and outputting the corrected fluorescence signal information, for example, displaying a fluorescence signal scattergram on a display according to the corrected fluorescence signal information, or transmitting the corrected fluorescence signal information to a remote server or a central station.

Of course, in some embodiments, after the correction, the fluorescence signal information before and after the correction may also be output, for example, the fluorescence signal scatter diagram after the correction is displayed on the display according to the fluorescence signal information after the correction, and the fluorescence signal scatter diagram before the correction is displayed on the display according to the fluorescence signal information before the correction, so as to facilitate the comparison of the user.

In some embodiments, after correction, a notification may also be issued that the fluorescent signal information has been corrected, such as by issuing a notification to the user via a display. In some embodiments, the processor 104 may also output the corrected magnitude of the fluorescence signal information, for example, by displaying the corrected magnitude of the fluorescence signal information in percentage form via a display. In some embodiments, after the correction, if the correction amplitude of the fluorescence signal information exceeds a set threshold, a notification that the fluorescence signal information has been corrected or an alarm is issued.

Example 3

Referring to fig. 5, the apparatus for correcting fluorescence signal information of a blood sample may include a memory 210 and a processor 230, where the memory 210 is used for storing a program, and the processor 230 is used for implementing the analysis method according to any one of the above embodiments by executing the program stored in the memory 210, for example, implementing the

above steps

300, 330 and 350, and also implementing step 370. The processor 230 may function similarly or identically to the processor 104 in the blood cell analyzer. This will be explained in detail below.

In one embodiment, processor 230 obtains fluorescence signal information of a blood sample treated with a fluorescence reagent, obtains a measured parameter of red blood cells in the blood sample, and modifies the fluorescence signal information according to the red blood cell parameter. The fluorescence signal mainly represents leukocyte granules or ghost granules, wherein the ghost granules refer to granules which remain the original shape and size of the cell membrane structure after the rupture of the plasma membrane after the hypotonic treatment of the red blood cells. In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.

As mentioned above, the data obtained by the processor 230 includes information of the fluorescence signal of the blood sample and the parameters of the red blood cells in the blood sample, and in one embodiment, the processor 230 can also obtain the forward scattered light signal information or the side scattered light signal information of the blood sample. These three types of data-first type of data: fluorescence information number information of blood sample, second type data: forward scattered light signal information or side scattered light signal information of the blood sample, and a third type of data: one or more of the parameters of the red blood cells in the blood sample may be transmitted by other devices to the processor 230.

Processor 230 modifies the fluorescence signal information according to the red blood cell parameters, which may include: and performing increase correction and/or decrease correction on the fluorescence signal information according to the erythrocyte parameters. The goal of the correction is: the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle no matter how the red blood cell parameters of HCT, RBC and HGB are changed.

(1) Augmentation correction

Processor 230 performs an augmentation correction on the fluorescence signal information according to the red blood cell parameters, including: and if the erythrocyte parameter is larger than a first preset value, performing augmentation correction on the fluorescence signal information. Further, in one embodiment, if the red blood cell parameter is greater than the first predetermined value, the larger the red blood cell parameter, the larger the magnitude of the increase correction performed by the processor 230 on the fluorescence signal information. The first preset value can be set as a reference according to the normal red blood cell parameter value under the condition of normal fluorescence intensity of the particles. As described above, the red blood cell parameters may include at least one of RBC, HGB, and HCT. When the red blood cell parameter includes only one of RBC, HGB, and HCT, then it is well understood that the red blood cell parameter is greater than the first preset value; when the red blood cell parameter includes two or three of RBC, HGB, and HCT, the red blood cell parameter being greater than the first preset value may refer to: and performing weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameters, wherein the specific weight value can be flexibly set in practice, the calculated value is used as the red blood cell parameter to be compared with a first preset value, and when the calculated value is larger than the first preset value, the red blood cell parameter is considered to be larger than the first preset value. For example, when the red blood cell parameters include three of RBC, HGB and HCT, the three parameters of RBC, HGB and HCT are calculated to perform weighted sum or weighted average calculation, the calculated value is regarded as the value of the red blood cell parameter to be compared with a first preset value, and when the calculated value is greater than the first preset value, the red blood cell parameter is greater than the first preset value.

(2) Reduction correction

Processor 230 applies a reduction correction to the fluorescence signal information based on the red blood cell parameters, including: and if the red blood cell parameter is smaller than a first preset value, performing reduction correction on the fluorescence signal information. Further, in one embodiment, if the red blood cell parameter is smaller than the first predetermined value, the smaller the red blood cell parameter, the larger the magnitude of the decrease correction of the fluorescence signal information by the processor 230

Processor 230 performs an increase correction and/or a decrease correction on the fluorescence signal information based on the red blood cell parameters, including: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information, wherein the correction function is an increasing function. In one embodiment, the modification function may be a linear function or a power function. In one embodiment, the modification function includes a first function and a second function, and both the first function and the second function are increasing functions; the processor substituting the red blood cell parameters and the fluorescence signal information as independent variables into a preset correction function comprises: substituting the erythrocyte parameter as an independent variable into the first function to obtain a correction coefficient, and substituting the correction coefficient as a constant and the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information. In one embodiment, the first function and the second function are both linear functions. For examples of the modification function, the first function and the second function, reference may be made to embodiment 1, or to specific embodiments which will be described in detail below, and details thereof are not repeated here.

In one embodiment of the processor 230, the corrected fluorescence signal information may also be output, such as displaying a fluorescence signal scatter plot on a display according to the corrected fluorescence signal information, or sending the corrected fluorescence signal information to a remote server, a central station.

Of course, in some embodiments, after the correction, the processor 230 may also output the fluorescence signal information before and after the correction, for example, display a fluorescence signal scatter diagram after the correction through the display according to the fluorescence signal information after the correction, and display a fluorescence signal scatter diagram before the correction through the display according to the fluorescence signal information before the correction, so as to facilitate the comparison by the user.

In some embodiments, after the correction, processor 230 may also issue a prompt to the user that the fluorescent signal information has been corrected, such as by issuing a prompt to the user via a display. In some embodiments, the processor 104 may also output the corrected magnitude of the fluorescence signal information, for example, by displaying the corrected magnitude of the fluorescence signal information in percentage form via a display. In some embodiments, after the correction, if the correction amplitude of the fluorescence signal information exceeds a set threshold, a notification that the fluorescence signal information has been corrected or an alarm is issued.

The present invention modifies the fluorescent signal information depending on one or more of the red blood cell parameters, such as HCT, RBC, and HGB. In the analysis of blood cells, red blood cell parameters such as HCT, RBC and HGB are conventional measurement parameters, so that the correction of fluorescence signal information can be completed under the conditions of not increasing reagent consumption and not increasing measurement channels, the measurement cost is not increased, and the accuracy of measurement results of measurement channels needing fluorescence signal information, such as NRBC channels, RET channels, DIFF channels and the like, is improved, for example, the results of WBC counting, NRBC classification, Baso classification and the like can be more accurately given.

The following is an example to illustrate how the present invention modifies the fluorescence signal information according to the red blood cell parameters.

In the following description of the relationship between fluorescence signal information and red blood cell parameters, the relationship between fluorescence signal of WBC particle clusters and red blood cell parameters in the experiment was mainly studied. The applicant counted 482 samples, and plotted scatter plots with the horizontal axis of HCT, RBC, and HGB and the vertical axis of WBC particle mass position in the fluorescence direction of these samples, as shown in fig. 6, 7, and 8.

As can be seen from the scatter diagrams of fig. 6 to 8, the position values of the WBC particle clusters in the fluorescence direction are inversely related to HCT, RBC, HGB, i.e., the smaller the HCT, RBC, HGB, the larger the position value of the WBC particle clusters in the fluorescence direction; the larger the HCT, RBC, HGB, the smaller the value of the WBC cluster position in the fluorescence direction.

One or more of the HCT, RBC, and HGB may be used to calculate a correction factor, and then the fluorescent signal information may be corrected using the calculated correction factor, for example, by multiplying the correction factor by the fluorescent signal information to obtain corrected fluorescent signal information. Taking fig. 6 to 8 as an example, the correction aims to make the position value of the WBC particle mass in the fluorescence direction substantially constant with the changes of the parameters HCT, RBC, HGB. In fig. 6 to 8, we can determine that the location value of the WBC particle mass in the fluorescence direction is equal to 2000 as the target horizontal line (i.e., the normal fluorescence signal intensity value of the leukocyte particle mass, which varies according to different systems), i.e., the location value of the WBC particle mass in the fluorescence direction is substantially maintained around 2000 regardless of the variation of the HCT, RBC, and HGB parameters. And with the change of the HCT, RBC and HGB parameters, the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle. In fig. 6, the first preset value for HCT is around 29; in fig. 7, the first preset value for RBC is around 2.3; in fig. 8, the HGB corresponds to a first preset value around 85. The first preset value can be set by taking the normal red blood cell parameter value under the condition of normal fluorescence intensity of the particle as reference, and can be understood as the red blood cell parameter value corresponding to the correction coefficient of 1.

Instead of taking the correction of fluorescence signal information in NRBC pass according to HCT as an example, the applicant constructed the first function f according to the fitting result of FIG. 6 described above₁₁(x₁)＝0.038x₁+0.09, wherein x₁Representing the value of HCT, and substituting the value of HCT into a correction coefficient which can be calculated; as shown in fig. 9, which is a function diagram of the first function, the abscissa is the value of HCT and the ordinate is the value of the correction coefficient, it can be seen that the larger HCT is, the larger the correction coefficient is, i.e., the first function is an increasing function. After calculating the correction coefficient, the fluorescence information signal is multiplied by the correction coefficient to obtain the corrected fluorescence signal information, such as the second function f₁₂(x_fl)＝k₁x_fl+ b, wherein k₁＝f₁₁(x₁)＝0.038x₁+0.09，x_flIndicating fluorescence signal information before correction; the correction function is therefore:

y_fl＝f₁(x₁,x_fl)＝(0.038x₁+0.09)*x_fl+b；

wherein y is_flRepresenting the corrected fluorescence signal information; b is a constant, e.g., 0; symbol is a multiplication number.

The first function used for the correction may correspond to a linear function, or may correspond to another functional form, such as a power function. In summary, the correction target is that the position value of the WBC particle mass in the fluorescence direction is substantially maintained near the target line, and the fluorescence signal value corresponding to the particle is corrected to be near the normal fluorescence signal intensity value of the particle, no matter how the parameters of HCT, RBC and HGB are changed.

As shown in fig. 10, a scatter plot of cell particles (primarily WBC particle mass in the upper right corner and ghost particle mass in the lower left corner) of three samples in NRBC channel is shown, where the abscissa is the value of the fluorescence signal information of the cell particles and the ordinate is the forward scatter signal information of the cell particles. The following is based on the above-mentioned correction function y_fl＝f₁(x₁,x_fl)＝(0.038x₁+0.09)*x_fl+ b the correction of the fluorescence signal information is performed for sample 1, sample 2 and sample 3, respectively, where b is taken to be 0.

In sample 1, RBC is 0.76 × 10 ^12/L, HGB is 26g/L, and HCT is 8.2%. As shown in FIG. 10(a), which is a scattergram of the unmodified cell particles obtained in the NRBC channel, the fluorescence signal of the cell particle mass can be seenThe number information is larger; the HCT value of sample 1 is 8.2%, which is substituted into the first function f₁₁(x₁)＝0.038x₁After +0.09, a correction factor of 0.4016 was obtained. The value of the fluorescence signal information of each cell particle in the sample 1 is multiplied by the correction coefficient 0.4016, and the corrected value of the fluorescence signal information of each cell particle is obtained, as shown in fig. 10 (d).

Sample 2 contains 3.25 × 10 ^12/L RBC, 71g/L HGB, and 23.8% HCT. As shown in fig. 10(b), which is a scattergram of unmodified cell particles obtained in the NRBC channel, it can be seen that fluorescence signal information of WBC particle clusters is moderate; the HCT value of sample 2 is 23.8%, which is substituted into the first function f₁₁(x₁)＝0.038x₁After +0.09, a correction factor of 0.9944 was obtained. The value of the fluorescence signal information of each cell particle in the sample 2 is multiplied by the correction coefficient 0.9944, and the corrected value of the fluorescence signal information of each cell particle can be obtained, as shown in fig. 10 (e).

Sample 3 contains RBC 7.04 × 10 ^12/L, HGB 158g/L, and HCT 50.3%. As shown in fig. 10(c), which is a scattergram of the cell particles before correction acquired in the NRBC channel, it can be seen that the fluorescence signal information of the cell particle mass is small; the HCT value of sample 3 is 50.3%, which is substituted into the first function f₁₁(x₁)＝0.038x₁After +0.09, a correction factor of 2.0014 was obtained. The value of the fluorescence signal information of each cell particle in the sample 3 is multiplied by the correction coefficient 2.0014, and the corrected value of the fluorescence signal information of each cell particle can be obtained, as shown in fig. 10 (f).

As can be seen from fig. 10(e) to 10(f), the fluorescence signals of the cell masses are corrected to a more appropriate range, i.e., a relatively eccentric position in the scattergram.

Because the channel dye methods such as NRBC channel, BASO channel, RET channel, DIFF channel and the like have the problem that the fluorescent signal of the leukocyte group is abnormal due to dye competition, the classification of the leukocyte group or NRBC group is easy to make mistakes. In leukocyte classification, the applicant has found through long-term development that in some samples, when cells are treated with a fluorescent dye and classified using a fluorescent signal and a scattered light signal, a case occurs in which the measurement result deviates from the result of microscopic examination. It has been found through intensive studies that the parameters of erythrocytes in such samples differ from normal values. Through verification, the red blood cell parameters and the fluorescence signal intensity have certain correlation, and the fluorescence signals can be corrected by using the red blood cell parameters. While not being bound by theory, applicants speculate that red blood cells may affect the staining behavior of other cells that need to be sorted, resulting in an abnormal fluorescence signal.

According to the embodiment of the application, the fluorescent signals are corrected according to the red blood cell parameters, so that the position of the white blood cell group in the scatter diagram is corrected, and the problem of inaccurate division of the white blood cell group is solved. In the classification of leukocytes, taking a fixed demarcation algorithm as an example, the embodiments of the present application can make a target particle group, such as a leukocyte group, or a nucleated erythrocyte group, or a reticulocyte group, be accurately located in a preset region of the fixed demarcation algorithm, thereby ensuring the accuracy of classification and counting. The method can be used for correcting the fluorescent signals by using the red blood cell parameters under the NRBC channel, the BASO channel, the WNB channel, the RET channel and the DIFF channel so as to correct the position of the white blood cell group in the scatter diagram.

Those skilled in the art will appreciate that all or part of the functions of the various methods in the above embodiments may be implemented by hardware, or may be implemented by computer programs. When all or part of the functions of the above embodiments are implemented by a computer program, the program may be stored in a computer-readable storage medium, and the storage medium may include: a read only memory, a random access memory, a magnetic disk, an optical disk, a hard disk, etc., and the program is executed by a computer to realize the above functions. For example, the program may be stored in a memory of the device, and when the program in the memory is executed by the processor, all or part of the functions described above may be implemented. In addition, when all or part of the functions in the above embodiments are implemented by a computer program, the program may be stored in a storage medium such as a server, another computer, a magnetic disk, an optical disk, a flash disk, or a removable hard disk, and may be downloaded or copied to a memory of a local device, or may be version-updated in a system of the local device, and when the program in the memory is executed by a processor, all or part of the functions in the above embodiments may be implemented.

The present invention has been described in terms of specific examples, which are provided to aid understanding of the invention and are not intended to be limiting. For a person skilled in the art to which the invention pertains, several simple deductions, modifications or substitutions may be made according to the idea of the invention.

Claims

1. A method of analyzing a blood sample, comprising:

obtaining a measured parameter of red blood cells in the blood sample;

2. The method for analyzing a blood sample according to claim 1, wherein the obtaining of fluorescence signal information of the blood sample treated with the fluorescent reagent comprises: and acquiring fluorescence signal information, forward scattered light signal information and/or side scattered light signal information of the blood sample treated by the fluorescent reagent.

3. The method for analyzing a blood sample according to claim 2, wherein the obtaining of fluorescence signal information of the blood sample treated with the fluorescent reagent comprises: and measuring the blood sample by adopting flow cytometry to acquire fluorescence signal information of the blood sample treated by the fluorescent reagent, and forward scattering light signal information and/or side scattering light signal information.

4. The method of analyzing a blood sample according to claim 2 or 3, further comprising: and carrying out white blood cell classification and/or counting according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

5. The method of analyzing a blood sample according to claim 2 or 3, further comprising: and counting the nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescence signal information.

6. The method of analyzing a blood sample according to claim 2 or 3, further comprising: and performing white blood cell counting and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescence signal information.

7. The method of analyzing a blood sample according to claim 2 or 3, further comprising: and performing basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

8. The method for analyzing a blood sample according to claim 1, further comprising, before acquiring fluorescence signal information of the blood sample treated with the fluorescent reagent: the method includes performing a red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, and then performing a hemolysis process and a fluorescent reagent process on the blood sample and measuring fluorescence signal information of the blood sample.

9. The method of claim 8, wherein said performing a red blood cell parameter measurement on said blood sample to determine said red blood cell parameter comprises: measuring a red blood cell parameter of the blood sample using a pinhole impedance method or an optical method to determine the red blood cell parameter.

10. The method of claim 1, wherein the modifying the fluorescence signal information according to the red blood cell parameter comprises: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as dependent variables.

11. The method of analyzing a blood sample according to claim 10, wherein the correction function is an increasing function of the dependent variable with respect to the independent variable.

12. The method of analyzing a blood sample according to claim 10, wherein the correction function is a linear function, a polynomial function, or a power function.

13. The method of claim 1, wherein the red blood cell parameter comprises at least one of hematocrit, red blood cell count, and hemoglobin concentration.

14. The method of claim 1, wherein after correcting the fluorescence signal information according to the red blood cell parameter, the method further comprises: and outputting the corrected fluorescence signal information.

15. The method of claim 14, wherein after correcting the fluorescence signal information according to the red blood cell parameter, further comprising: and outputting the fluorescence signal information before and after correction.

16. The method of claim 1, wherein after correcting the fluorescence signal information according to the red blood cell parameter, the method further comprises: and sending out a prompt that the fluorescence signal information is corrected or sending out an alarm.

17. The method of analyzing a blood sample according to claim 16, wherein the indicating that the information on the emitted fluorescent signal has been corrected comprises: and outputting the corrected amplitude of the fluorescence signal information.

18. The method of claim 16, wherein after correcting the fluorescence signal information according to the red blood cell parameter, further comprising: and if the correction amplitude of the fluorescence signal information exceeds a set threshold value, sending out a prompt that the fluorescence signal information is corrected or sending out an alarm.

19. A blood cell analyzer, comprising:

20. The blood cell analyzer of claim 19, wherein the processor obtains fluorescence signal information of the blood sample treated with the fluorescent reagent, and forward scattered light signal information and/or side scattered light signal information of leukocyte particles.

21. The blood cell analyzer of claim 20, wherein the optical detection device comprises a light source and a flow chamber having an aperture, leukocyte particles in the blood sample treated with the fluorescent reagent flow in the flow chamber and pass through the aperture one by one, and light emitted from the light source irradiates the particles in the aperture and generates a fluorescent signal and a scattered light signal, and outputs information of the fluorescent signal, and information of a forward scattered light signal and/or information of a side scattered light signal to the processor after conversion by the optical detection device.

22. The blood cell analyzer of claim 20 or 21, wherein the processor further performs leukocyte classification and/or enumeration based on the side scattered light values and the corrected fluorescence signal information of the blood sample.

23. The blood cell analyzer of claim 20 or 21, wherein the processor further performs a nucleated red blood cell count based on the forward scattered light value of the blood sample and the corrected fluorescence signal information.

24. The blood cell analyzer of claim 20 or 21, wherein the processor further performs white blood cell counting and/or reticulocyte recognition and/or basophil classification based on the forward scattered light value of the blood sample and the corrected fluorescence signal information.

25. The blood cell analyzer of claim 20 or 21, wherein the processor further performs basophil classification based on the side scattered light value of the blood sample and the corrected fluorescence signal information.

26. The blood cell analyzer of claim 19, wherein the reaction chamber provides a reaction site for the blood sample and a reagent required for measuring a parameter of red blood cells, the optical detection device measures the parameter of red blood cells of the blood sample treated by the reagent and outputs optical signal information, and the processor determines the parameter of red blood cells according to the optical signal information; the reaction cell carries out hemolysis treatment and fluorescence reagent treatment on the blood sample, then the optical detection device measures the treated blood sample and outputs the fluorescence signal information of the blood sample.

27. The blood cell analyzer of claim 19, wherein the processor is configured to: and substituting the erythrocyte parameters and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as dependent variables.

28. The blood cell analyzer of claim 27, wherein the correction function is an increasing function of the dependent variable relative to the independent variable.

29. The hematology analyzer of claim 19, wherein the red blood cell parameters include at least one of hematocrit, red blood cell count, and hemoglobin concentration.

30. The blood cell analyzer of claim 19, wherein the processor further outputs the corrected fluorescence signal information.

31. A computer-readable storage medium, in which a program is stored, the program being executable by a processor to implement the method of any one of claims 1 to 18.