CN110887818B - Analysis method of blood sample, blood cell analyzer and storage medium - Google Patents

Abstract

A blood sample analysis method, a blood cell analyzer and a storage medium acquire fluorescence signal information of a blood sample treated by a fluorescent reagent; obtaining measured parameters of red blood cells in the blood sample; and correcting the fluorescent signal information and/or sending out abnormal prompt of the fluorescent signal information according to the red blood cell parameters.

Description

A blood sample analysis method, blood cell analyzer and storage medium

technical field

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

Background technique

A blood cell analyzer is an instrument that can detect cells in blood, for example, it can classify and count white blood cells, red blood cells, platelets, nucleated red blood cells, reticulocytes and other cells in blood.

Taking white blood cell detection as an example, the most common method is the laser light scattering method, that is, blood cell analyzers use scattered light to classify and count white blood cells. Scattered light includes three types of light signals: forward scattered light, side scattered light, and fluorescent signal. 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 intracellular DNA, RNA and other substances that can be dyed by fluorescent dyes. The white blood cells can be classified by using these light signals, and the count value of the white blood cells can be obtained at the same time.

When applying the existing white blood cell differential counting method to do some blood tests, the accuracy can not meet the requirements. Therefore, it is necessary to develop a leukocyte classification method with better accuracy.

Contents of the invention

The application provides an analysis method of a blood sample, a blood cell analyzer and a storage medium.

An analysis method for a blood sample is provided, comprising:

Obtain the fluorescence signal information of the blood sample treated with the fluorescent reagent;

obtaining the measured red blood cell parameters in the blood sample;

The fluorescent signal information is corrected according to the red blood cell parameters and/or an abnormality prompt is issued for the fluorescent signal information.

In one of the embodiments, the acquiring the fluorescent signal information of the blood sample treated with the fluorescent reagent includes: acquiring the fluorescent signal information of the blood sample treated with the fluorescent reagent, and the forward scattered light signal information and/or the lateral Scattered light signal information.

In one of the embodiments, the acquiring the fluorescent signal information of the blood sample treated with the fluorescent reagent comprises: measuring the blood sample by flow cytometry to acquire the fluorescent signal information of the blood sample treated with the fluorescent reagent , and forward scattered light signal information and/or side scattered light signal information.

In one embodiment, further comprising: performing white blood cell classification and/or counting according to the side scattered light value of the blood sample and the corrected fluorescent signal information.

In one of the embodiments, the method further includes: counting nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescent signal information.

In one embodiment, further comprising: performing white blood cell count and/or reticulocyte identification and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescent signal information.

In one of the embodiments, the method further includes: classifying basophils according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

In one of the embodiments, before obtaining the fluorescence signal information of the blood sample treated with the fluorescent reagent, it also includes: performing red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, and then performing hemolysis on the blood sample and fluorescent reagents, and measure the fluorescent signal information of the blood sample.

In one embodiment, the measuring the red blood cell parameters of the blood sample to determine the red blood cell parameters includes: measuring the red blood cell parameters of the blood sample by using a small hole impedance method or an optical method to determine the red blood cell parameters .

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

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

In one of the embodiments, the correction function is a linear function, a polynomial function or a power function.

In one of the embodiments, the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration.

In one of the embodiments, after correcting the fluorescent signal information according to the red blood cell parameters, the method further includes: outputting the corrected fluorescent signal information.

In one of the embodiments, after correcting the fluorescent signal information according to the red blood cell parameters, the method further includes: outputting the fluorescent signal information before and after correction.

In one of the embodiments, after correcting the fluorescent signal information according to the red blood cell parameters, it further includes: issuing a prompt or issuing an alarm that the fluorescent signal information has been corrected.

In one of the embodiments, the prompting that the fluorescent signal information has been corrected includes: outputting a correction range of the fluorescent signal information.

In one of the embodiments, after correcting the fluorescent signal information according to the red blood cell parameters, it further includes: if the correction range of the fluorescent signal information exceeds the set threshold, sending a prompt or issuing a message that the fluorescent signal information has been corrected. Call the police.

Also provided is a hematology analyzer comprising:

At least one reaction pool is used to provide a reaction place for blood samples and reagents;

The optical detection device is used to irradiate the blood sample treated with the reagent with light, collect the optical signal generated by each particle in the blood sample treated with the reagent due to the light irradiation, and convert it into an electrical signal to output the optical signal information;

A delivery device, used to deliver the reagent-treated blood sample in the reaction cell to the optical detection device;

a processor, configured to receive and process the optical signal information output by the optical detection device to obtain the measurement parameters of the blood sample; Red blood cell parameters in the blood sample, correcting the fluorescent signal information according to the red blood cell parameters and/or sending out abnormal fluorescent signal information prompts.

In one of the embodiments, the processor acquires the fluorescent signal information of the blood sample treated with the fluorescent reagent, and the forward scattered light signal information and/or the side scattered light signal information of the white blood cell particles.

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

In one of the embodiments, the processor further classifies and/or counts white blood cells according to the side scattered light value of the blood sample and the corrected fluorescent signal information.

In one of the embodiments, the processor also counts nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescent signal information.

In one of the embodiments, the processor further performs white blood cell count and/or reticulocyte identification and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescent signal information.

In one of the embodiments, the processor further classifies basophils according to the side scattered light value of the blood sample and the corrected fluorescence signal information.

In one of the embodiments, the reaction pool provides a reaction place for the blood sample and the reagents required for the measurement of red blood cell parameters, and the optical detection device measures the red blood cell parameters of the blood sample processed by the reagents, and outputs Optical signal information, the processor determines the red blood cell parameters according to the optical signal information; the reaction pool performs hemolysis treatment and fluorescent reagent treatment on the blood sample, and then the optical detection device measures the processed blood sample , outputting the fluorescence signal information of the blood sample.

In one embodiment, the processor is configured to: substitute the red blood cell parameters and the fluorescent signal information as independent variables into a preset correction function, so as to obtain corrected fluorescent signal information as a dependent variable.

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

In one of the embodiments, the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration.

In one of the embodiments, the processor also outputs the corrected fluorescent signal information.

A computer-readable storage medium stores a program, and the program can be executed by a processor to implement the method described in any one of the above embodiments.

According to the blood sample analysis method, the blood cell analyzer and the computer-readable storage medium of the above-mentioned embodiments, the red blood cell parameters are used to correct the fluorescent signal information, thereby improving the accuracy of the fluorescent signal information.

Description of drawings

Fig. 1 is a schematic structural diagram of a blood cell analyzer of an embodiment;

Fig. 2 is a flowchart of an analysis method of a blood sample in an embodiment;

Fig. 3 is a flow chart of another embodiment of a blood sample analysis method;

Fig. 4 is a flowchart of a blood sample analysis method in another embodiment;

Fig. 5 is a schematic structural diagram of a device for correcting fluorescence signal information of a blood sample according to an embodiment;

Fig. 6 is a scatter diagram of the position value and HCT of the WBC particle cluster in the fluorescence direction in statistics of multiple samples;

Fig. 7 is a scatter diagram of the position value of the WBC particle cluster in the fluorescence direction and the RBC in the statistics of multiple samples;

Figure 8 is a scatter diagram of the position value of the WBC particle cluster in the fluorescence direction and the HGB in the statistics of multiple samples;

FIG. 9 is a function diagram of a first function corrected using HCT in an embodiment;

Fig. 10 is a schematic diagram of fluorescence signal information of cell particles before and after correction in three samples.

Detailed ways

The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings. Wherein, similar elements in different implementations adopt associated similar element numbers. In the following implementation manners, many details are described for better understanding of the present application. However, those skilled in the art can readily recognize that some of the features can be omitted in different situations, or can be replaced by other elements, materials, and methods. In some cases, some operations related to the application are not shown or described in the description, this is to avoid the core part of the application being overwhelmed by too many descriptions, and for those skilled in the art, it is necessary to describe these operations in detail Relevant operations are not necessary, and they can fully understand the relevant operations according to the description in the specification and general technical knowledge in the field.

In addition, the characteristics, operations or characteristics described in the specification can be combined in any appropriate manner to form various embodiments. At the same time, the steps or actions in the method description can also be exchanged or adjusted in a manner obvious to those skilled in the art. Therefore, various sequences in the specification and drawings are only for clearly describing a certain embodiment, and do not mean a necessary sequence, unless otherwise stated that a certain sequence must be followed.

The serial numbers assigned to components in this document, such as "first", "second", etc., are only used to distinguish the described objects, and do not have any sequence or technical meaning. The "connection" and "connection" mentioned in this application all include direct and indirect connection (connection) unless otherwise specified.

Taking the common five-differentiation blood cell analyzer as an example, the equipment generally has a channel that requires fluorescence signal measurement. It uses three kinds of scattered light signals, such as forward scattered light, side scattered light, and fluorescent signal, to classify and count cells, for example White blood cell count, nucleated red blood cell classification, and some basophil classification can be performed.

Some English nouns in this application are 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 corpuscular volume;

WBC: white blood cells (number);

BASO: Basophils (number).

Common five-differentiation hematology analyzers have channels that require fluorescent signal measurement, which use three optical signals, including forward scattered light, side scattered light, and side fluorescent signal, to classify and count cells, for example, white blood cell counts can be performed , Classification of nucleated red blood cells, and some can also classify basophils. The channel mentioned in this article refers to the 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, and 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. What can be known will not be repeated here.

Classification and counting are carried out by obtaining the optical signal of the cells, and when the WBC particle clusters of different samples appear in different positions, the classification will be wrong. In the classification of white blood cells, the applicant has discovered through long-term research and development that, in some samples, when the cells are treated with fluorescent dyes and the cells are classified using fluorescent signals and scattered light signals, there will be deviations between the measurement results and the results of microscopic examination . Through in-depth research, it was found that in such samples, the parameters of red blood cells were different from normal values. Through verification, it is found that there is a certain correlation between the red blood cell parameters and the intensity of the fluorescent signal, and the red blood cell parameters can be used to correct the fluorescent signal. While not being bound by theory, the applicant speculates that red blood cells may affect the staining behavior of other cells that need to be sorted, resulting in abnormal fluorescent signals.

The applicant found in practice that in the currently common five-differentiation blood cell analyzer, we might as well take the NRBC channel as an example. When the cells are stained with fluorescent dyes, the positions of the cell fluorescence signals in the fluorescence direction of different samples will appear. In the process of counting and classifying cells (such as white blood cells), if the fluorescent signal is too large or too small, it will affect the algorithm of counting and classifying white blood cells (for example, the fluorescent signal affects the setting of the gate), which will affect the counting and counting of NRBC channels. Classification will further increase the risk of clinical misdiagnosis. Let’s take the RET channel again as an example. During the testing process, different distributions of red blood cells will result in different processing degrees of fluorescent dyes on the cells, resulting in different signals of cells in the direction of fluorescence, which will affect the classification of red blood cells such as the recognition of RET.

Therefore, for those channels that use fluorescent dyes, such as NRBC channels, RET channels, and DIFF channels, when these channels are used for sample measurement, if the fluorescence signal correction is not performed, the fluorescence signal of WBC particles will appear too large or too small, In the case of a limited number of bytes in the signal collector, due to the maximum and minimum limits of the signal, the WBC particle cluster will eventually appear too far to the left or right in the scatter diagram, resulting in distortion of the channel scatter diagram , the lack of necessary scattergram information makes it difficult for subsequent classification, and it is easy to make mistakes in the classification of blood ghosts, nucleated red blood cells, and basophils, thus giving wrong counting and classification results.

To sum up, for those channels using fluorescent dyes, such as NRBC channel, BASO channel, WNB channel, RET channel and DIFF channel, etc., there is a need for a technical solution to correct the difference in fluorescence signals of different samples in these channels.

In response to the above problems, the applicant discovered after theoretical analysis of reagent action and research on a large number of samples that for channels using fluorescent dyes—for example, NRBC channels, during the actual measurement process, the fluorescent signal of its WBC particles is related to red blood cell count (RBC), erythrocyte hemoglobin Content (HGB) and hematocrit (HCT) are negatively correlated, that is, the same sample or blood sample, the smaller the RBC, HGB, and HCT, the larger the fluorescence signal of WBC particles; the larger the RBC, HGB, and HCT, the larger the WBC particle The fluorescent signal is smaller. Therefore, RBC, HGB, and HCT can be used to correct the fluorescent signal of WBC particles and/or send out abnormal fluorescent signal information, that is, the fluorescent signal information can be corrected according to red blood cell parameters and/or send out abnormal fluorescent signal information. It can only be corrected, or only an abnormal prompt can be issued, and of course, both correction and abnormal prompt can be issued.

Example 1

A blood cell analyzer according to an embodiment of the present invention may mainly include the structure shown in FIG. 1 : at least one reaction cell 101 , an optical detection device 102 , a delivery device 103 and a processor 104 , which will be described in detail below.

The reaction pool 101 is used to provide a reaction place for blood samples and reagents to be prepared into sample liquids. Specifically, the blood sample obtained from blood collection can be diluted and labeled with a fluorescent staining reagent to obtain a sample solution. Commonly used fluorescent staining reagents can be pyronine, acridine orange and thiazole orange, etc.

The optical detection device 102 is used to irradiate the reagent-treated blood sample, that is, the above-mentioned sample liquid, collect the optical signal generated by each particle in the reagent-treated blood sample due to light irradiation, and convert it into an electrical signal , to output optical signal information. The optical signal here may be a forward scattered light signal, a side scattered light signal, or a fluorescent signal. In one embodiment of the optical detection device 102, it can include a light source 1021 and a sheath flow chamber 1022 with an orifice 10221, etc., particles in the blood sample can flow in the sheath flow chamber 1022 and pass through the orifice 10221 one by one, The light emitted by the light source 1021 can irradiate the particles in the aperture 10221 and correspondingly generate scattered light signals and/or fluorescence signals. The optical detection device 102 can also include lens groups 1023, photoelectric sensors 1024 (such as photodiodes, photomultiplier tubes, etc.) An element is formed in the processor 104 or separately, so that the lens group 1023 can capture the corresponding scattered light signal and fluorescent signal, and the photoelectric sensor 1024 can convert the captured optical signal (refers to the scattered light signal and fluorescent signal, etc.) into an electrical signal, Then the A/D converter converts the electrical signal through A/D to obtain a digital signal, which can be output as an optical signal information.

The delivery device 103 is used to deliver the blood sample in the reaction cell 101 , that is, the sample liquid, which has been treated with reagents, to the optical detection device 102 .

The processor 104 is configured to receive and process the optical signal information output by the optical detection device 102 to obtain measurement parameters of the blood sample. In one embodiment of the processor 104, it acquires the fluorescence signal information of the blood sample treated with the fluorescent reagent, acquires the measured red blood cell parameters in the blood sample, and corrects the fluorescence signal information according to the red blood cell parameters. The particles represented by the fluorescent signal are mainly leukocyte particles or ghost particles, where the ghost particles refer to the particles that maintain the original shape and size of the cell membrane structure after the plasma membrane ruptures after the red blood cells are treated with hypotonicity. In one embodiment, the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration.

As mentioned above, the data acquired by the processor 104 includes the fluorescence signal information of the blood sample and the red blood cell parameters in the blood sample. In an embodiment, the processor 104 can also acquire the forward scattered light signal information or Side scattered light signal information. These three types of data—that is, the first type of data: the fluorescence information number information of the blood sample, the second type of data: the forward scattered light signal information or the side scattered light signal information of the blood sample, and the third type of data: all One or more of the red blood cell parameters in the blood sample——can be transmitted to the processor in the blood cell analyzer of the present invention by other equipment, and can also be measured by the blood cell analyzer of the present invention to obtain, as illustrated below .

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

Acquisition of the second type of data (forward scattered light signal information or side scattered light signal information of the blood sample) by the processor 104: the forward scattered light signal information and the side scattered light signal information may also be obtained when obtaining the fluorescence signal Optical signal information; for example, the reaction pool 101 performs hemolysis treatment and fluorescent reagent treatment on the blood sample, and the delivery device 103 transports the blood sample after hemolysis treatment and fluorescent reagent treatment to the optical detection device 102, and the optical detection device 102 performs the hemolysis treatment and fluorescent reagent treatment. The blood sample treated with the fluorescent reagent is measured, and one or both of the fluorescent signal information, the forward scattered light signal information and the side scattered light signal information of the blood sample are output to the processor 104 . Specifically, through the delivery of the delivery device 103, the cell particles in the blood sample treated with the fluorescent reagent flow in the sheath flow chamber 1022, and pass through the orifice 10221 one by one, and the light emitted by the light source 1021 irradiates the particles in the orifice 10221. The cell particles correspondingly generate the fluorescence signal and the scattered light signal of the cell particle (referring to one or both of the forward scattered light signal information and the side scattered light signal information), and are converted by the optical detection device 10221 to the processor 104 outputs fluorescence signal information, and forward scattered light signal information or side scattered light signal information.

The processor 104 can perform the following cell detection through the acquired fluorescence signal information and scattered light signal information. The white blood cell classification and/or counting is performed according to the side scattered light value of the blood sample and the corrected fluorescent signal information, for example, four classifications of the white blood cells are performed through the DIFF channel. Nucleated red blood cell count is performed according to the forward scattered light value of the blood sample and the corrected fluorescent signal information, for example, the nucleated red blood cell count is performed through the NRBC channel. Perform white blood cell count and/or reticulocyte identification and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescent signal information, for example, perform reticulocyte classification and/or count through the RET channel. Basophil classification is performed based on the side scatter light value of the blood sample and the corrected fluorescence signal information, such as basophil classification by the BASO channel.

Acquisition of the third type of data (red blood cell parameters in the blood sample) by the processor 104: the reaction pool 101 provides a reaction site for the blood sample and the reagents required for red blood cell parameter measurement, and the delivery device 103 transfers the blood after the reagent treatment The sample is sent to the optical detection device 102, the optical detection device 102 measures the red blood cell parameters of the blood sample processed by the reagent, and outputs optical signal information, and the processor 104 determines the red blood cell parameters according to the optical signal information. That is, the red blood cell parameters in the blood sample can be obtained through optical detection. Of course, in other embodiments, the red blood cell parameters in the blood sample can also be obtained by measuring the small hole impedance method, for example, measuring the blood sample by the small hole impedance method from the detection device of the blood cell analyzer itself or other instruments.

The above is the description of the acquisition of the above three types of data by the processor 104, and the modification of the processor 104 will be described below.

In an embodiment, the processor 104 correcting the fluorescence signal information according to the red blood cell parameters may include: performing increase correction and/or decrease correction on the fluorescence signal information according to the red blood cell parameters. The goal of the correction is: no matter how the red blood cell parameters such as HCT, RBC, and HGB change, the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle.

(1) Increase correction

The processor 104 performing increasing correction on the fluorescent signal information according to the red blood cell parameter includes: performing increasing correction on the fluorescent signal information if the red blood cell parameter is greater than a first preset value. 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 magnitude of the increase correction performed by the processor 104 on the fluorescent signal information is. The setting of the first preset value can be based on the parameter value of normal red blood cells under the condition of normal fluorescence intensity of the particles as a reference.

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

(2) Reduce correction

The processor 104 performing reduction correction on the fluorescence signal information according to the red blood cell parameter includes: performing reduction correction on the fluorescence signal information if the red blood cell parameter is smaller than a first preset value. Further, in one embodiment, if the red blood cell parameter is smaller than the first preset value, the smaller the red blood cell parameter is, the larger the reduction and correction of the fluorescent signal information by the processor 104 will be.

As mentioned above, the red blood cell parameters may include at least one of RBC, HGB and HCT. When the red blood cell parameters include only one of RBC, HGB and HCT, it is easy to understand that the red blood cell parameters are less than the first preset value; when the red blood cell parameters include two or three of RBC, HGB and HCT, the red blood cell parameters are less than The first preset value may refer to: performing a weighted sum of two or three of RBC, HGB, and HCT included in the red blood cell parameters, or a weighted average calculation. The specific weight value can be flexibly set in practice, and the calculated The value of is used as the red blood cell parameter to compare with the first preset value, and when the calculated value is smaller than the first preset value, it is considered that the red blood cell parameter is smaller than the first preset value. For example, when the red blood cell parameters include RBC, HGB and HCT, the weighted sum of the three parameters of RBC, HGB and HCT is calculated, or the weighted average is calculated, and the calculated value is regarded as the value of the red blood cell parameter to be compared with the first Compared with the preset value, when the calculated value is smaller than the first preset value, it means that the red blood cell parameter is smaller than the first preset value.

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

In an embodiment, the correction function may be a linear function or a power function. In one embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor uses the red blood cell parameters and the fluorescent signal information as an automatic Substituting variables into the preset correction function includes: substituting the red blood cell parameters as an independent variable into the first function to obtain a correction coefficient, using the correction coefficient as a constant and the fluorescent signal information as an independent variable into the second function In order to obtain the corrected fluorescent signal information as the dependent variable. In an embodiment of the processor 104, it can also output the corrected fluorescence signal information, for example, display the fluorescence signal scatter diagram on the display according to the corrected fluorescence signal information, or send the corrected fluorescence signal information to the remote server , Central Station. Of course, in some embodiments, the fluorescence signal information before and after correction can also be output, for example, the scatter diagram of the fluorescence signal after correction can be displayed on the display according to the fluorescence signal information after correction, and the fluorescence signal information before correction can be displayed according to the fluorescence signal information before correction. The scatter diagram of the fluorescent signal before correction is displayed on the display to facilitate user comparison.

In some embodiments, after correction, the processor 104 may also send out a prompt or alarm that the fluorescent signal information has been corrected, for example, send a prompt to the user through a display. In some embodiments, the processor 104 may also output the correction range of the fluorescence signal information, for example, display the correction range of the fluorescence signal information in a percentage form through a display. In some embodiments, if the correction range of the fluorescent signal information exceeds a set threshold, a prompt or an alarm that the fluorescent signal information has been corrected is issued.

The correction function, the first function, and the second function are illustrated below as examples.

In the following formulas (1) to (7), y _fl represents the fluorescent signal information after correction, x _fl represents the fluorescent signal information before correction, and x ₁ , x ₂ , and x ₃ represent the values of HCT, RBC, and HGB, respectively .

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

y _fl =f ₁ (x ₁ ,x _fl );

Further, the first function included in the correction function f ₁ (x ₁ , x _fl ) may be f ₁₁ (x ₁ ), and the second function may be f ₁₂ (x _fl ), for example, the second function f ₁₂ (x _fl ) =k ₁ x _fl +b, k ₁ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₁ =f ₁₁ (x ₁ ).

(2) When the red blood cell parameters only include RBC, the correction function can have the following form:

y _fl =f ₂ (x ₂ ,x _fl );

Further, the first function included in the correction function f ₂ (x ₂ , x _fl ) may be f ₂₁ (x ₂ ), and the second function may be f ₂₂ (x _fl ), for example, the second function f ₂₂ (x _fl ) =k ₂ x _fl +b, k ₂ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₂ =f ₂₁ (x ₂ ).

(3) When the red blood cell parameters only include HGB, the correction function can have the following form:

y _fl =f ₃ (x ₃ ,x _fl );

Further, the first function included in the correction function f ₃ (x ₃ , x _fl ) may be f ₃₁ (x ₃ ), and the second function may be f ₃₂ (x _fl ), for example, the second function f ₃₂ (x _fl ) =k ₃ x _fl +b, k ₃ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₃ =f ₃₁ (x ₃ ).

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

y _fl =f ₄ (x ₁ ,x ₂ ,x _fl );

Further, the first function included in the correction function f ₄ (x ₁ , x ₂ , x _fl ) may be f ₄₁ (x ₁ , x ₂ ), and the second function may be f ₄₂ (x _fl ), for example, the second function f ₄₂ (x _fl )=k ₄ x _fl +b, k ₄ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₄ =f ₄₁ (x ₁ ,x ₂ ).

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

y _fl =f ₅ (x ₂ ,x ₃ ,x _fl );

Further, the first function included in the correction function f ₅ (x ₂ , x ₃ , x _fl ) may be f ₅₁ (x ₂ , x ₃ ), and the second function may be f ₅₂ (x _fl ), for example, the second function f ₅₂ (x _fl )=k ₅ x _fl +b, k ₅ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₅ =f ₅₁ (x ₂ ,x ₃ ).

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

y _fl =f ₆ (x ₁ ,x ₃ ,x _fl );

Further, the first function included in the correction function f ₆ (x ₁ , x ₃ , x _fl ) may be f ₆₁ (x ₁ , x ₃ ), and the second function may be f ₆₂ (x _fl ), for example, the second function f ₆₂ (x _fl )=k ₆ x _fl +b, k ₆ and b are constants, and the above-mentioned substitution of the correction coefficient as a constant into the second function may refer to k ₆ =f ₆₁ (x ₁ ,x ₃ ).

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

y _fl =f ₇ (x ₁ ,x ₂ ,x ₃ ,x _fl );

Further, the first function included in the correction function f ₇ (x ₁ , x ₂ , x ₃ , x _fl ) may be f ₇₁ (x ₁ , x ₂ , x ₃ ), and the second function may be f ₇₂ (x _fl ), for example, the second function f ₇₂ (x _fl )=k ₇ x _fl +b, k ₇ and b are constants, and the above-mentioned substitution of the correction coefficient into the second function as a constant may refer to k ₇ =f ₇₁ (x ₁ ,x ₂ ,x ₃ ).

Example 2

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

Step 310: Obtain fluorescence signal information of the blood sample treated with the fluorescent reagent. In an embodiment, step 310 may include: acquiring fluorescence signal information of the blood sample treated with the fluorescent reagent, and forward scattered light signal information or side scattered light signal information.

Step 330: Obtain the measured red blood cell parameters in the blood sample. In one embodiment, the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration.

In the above steps, the data of fluorescence signal information, forward scattered light signal information and side scattered light signal information obtained in step 310 can be measured in real time or non-real time, for example, these data are stored in a memory , step 310 fetches data from the memory. Similarly, the red blood cell parameters obtained in step 330 can 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 is obtained from the memory. Red blood cell parameters in the blood sample are acquired.

Let’s take the real-time measurement of the fluorescence signal information of the blood sample and the red blood cell parameters in the blood sample as an example. Please refer to FIG. 3 . An embodiment of the analysis method may also include steps 300 and 320, which will be described in detail below.

Step 300: Perform hemolysis treatment and fluorescent reagent treatment on the blood sample, and measure the fluorescent signal information of the blood sample.

Step 320: Perform red blood cell parameter measurement on the blood sample to determine the red blood cell parameter. In one embodiment, if the analysis method has step 300 and step 320 at the same time, step 320 may be performed first, and then step 300 is performed, for example, red blood cell parameters are measured on the blood sample to determine the red blood cell parameters, and then the The blood sample is subjected to hemolysis treatment and fluorescent reagent treatment, and the fluorescence signal information of the blood sample is measured.

It may be advisable to obtain the fluorescence signal information and scattered light signal information (forward scattered light signal information or side scattered light signal information) of the blood sample by real-time measurement. In an embodiment of the analysis method, step 310 may be to use flow cytometry to analyze The blood sample is measured to obtain the fluorescence signal information of the blood sample treated with the fluorescent reagent, and the forward scattered light signal information or the side scattered light signal information.

Through the obtained fluorescent signal information and scattered light signal information, the following cell detection can be performed. The white blood cell classification and/or counting is performed according to the side scattered light value of the blood sample and the corrected fluorescent signal information, for example, four classifications of the white blood cells are performed through the DIFF channel. Nucleated red blood cell count is performed according to the forward scattered light value of the blood sample and the corrected fluorescent signal information, for example, the nucleated red blood cell count is performed through the NRBC channel. Perform white blood cell count and/or reticulocyte identification and/or basophil classification according to the forward scattered light value of the blood sample and the corrected fluorescent signal information, for example, perform reticulocyte classification and/or count through the RET channel. Basophil classification is performed based on the side scatter light value of the blood sample and the corrected fluorescence signal information, such as basophil classification by the BASO channel.

Step 350: Correcting the fluorescent signal information according to the red blood cell parameters and/or issuing a prompt for abnormal fluorescent signal information. That is, the fluorescent signal information can be corrected according to the red blood cell parameters and/or an abnormality prompt can be issued for the fluorescent signal information. It can only be corrected, or only an abnormal prompt can be issued, of course, it can also be corrected and an abnormal prompt can be issued

In some embodiments, step 350 correcting the fluorescent signal information according to the red blood cell parameters includes: performing an increase correction and/or a decrease correction on the fluorescent signal information according to the red blood cell parameters. The goal of the correction is: no matter how the red blood cell parameters such as HCT, RBC, and HGB change, the fluorescent signal value corresponding to the particle is corrected to be close to the normal fluorescent signal intensity value of the particle.

(1) Increase correction

Step 350 performing increasing correction on the fluorescent signal information according to the red blood cell parameter includes: performing increasing correction on the fluorescent signal information if the red blood cell parameter is greater than a first preset value. Further, in one 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 magnitude of the increase correction performed on the fluorescent signal information in step 350 is. The setting of the first preset value can be based on the parameter value of normal red blood cells under the condition of normal fluorescence intensity of the particles as a reference.

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

(2) Reduce correction

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

As mentioned above, the red blood cell parameters may include at least one of RBC, HGB and HCT. When the red blood cell parameters include only one of RBC, HGB and HCT, it is easy to understand that the red blood cell parameters are less than the first preset value; when the red blood cell parameters include two or three of RBC, HGB and HCT, the red blood cell parameters are less than The first preset value may refer to: performing a weighted sum of two or three of RBC, HGB, and HCT included in the red blood cell parameters, or a weighted average calculation. The specific weight value can be flexibly set in practice, and the calculated The value of is used as the red blood cell parameter to compare with the first preset value, and when the calculated value is smaller than the first preset value, it is considered that the red blood cell parameter is smaller than the first preset value. For example, when the red blood cell parameters include RBC, HGB and HCT, the weighted sum of the three parameters of RBC, HGB and HCT is calculated, or the weighted average is calculated, and the calculated value is regarded as the value of the red blood cell parameter to be compared with the first Compared with the preset value, when the calculated value is smaller than the first preset value, it means that the red blood cell parameter is smaller than the first preset value.

Step 350 performs increase correction and/or decrease correction on the fluorescent signal information according to the red blood cell parameters, including: substituting the red blood cell parameters and the fluorescent signal information as independent variables into a preset correction function to obtain the corrected Fluorescent signal information of , wherein the correction function is an increasing function. In one embodiment, the correction function is a linear function or a power function. In one embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor uses the red blood cell parameters and the fluorescent signal information as an automatic Substituting variables into the preset correction function includes: substituting the red blood cell parameters as an independent variable into the first function to obtain a correction coefficient, using the correction coefficient as a constant and the fluorescent signal information as an independent variable into the second function In order to obtain the corrected fluorescent signal information. In an embodiment, both the first function and the second function are linear functions. For examples of the correction function, the first function and the second function, reference may be made to Embodiment 1, which will also be described in detail in the following embodiments, and will not be repeated here.

Please refer to FIG. 4 , an embodiment of the analysis method may also include step 370: outputting the corrected fluorescence signal information, for example, displaying the fluorescence signal scattergram on the display according to the corrected fluorescence signal information, or displaying the corrected fluorescence signal information The signal information is sent to the remote server and the central station.

Certainly, in some embodiments, after correction, the fluorescence signal information before correction and after correction can also be output, for example, displaying the corrected fluorescence signal scatter diagram on the display according to the corrected fluorescence signal information, and displaying the corrected Fluorescent signal information displays the scatter diagram of the fluorescent signal before correction through the monitor, which is convenient for users to compare.

In some embodiments, after correction, a reminder that the fluorescent signal information has been corrected may also be issued, for example, a reminder is issued to the user through a display. In some embodiments, the processor 104 may also output the correction range of the fluorescence signal information, for example, display the correction range of the fluorescence signal information in a percentage form through a display. In some embodiments, after correction, if the correction magnitude of the fluorescent signal information exceeds a set threshold, a prompt or an alarm is sent out that the fluorescent signal information has been corrected.

Example 3

Corresponding to the above method for correcting fluorescence signal information of blood samples, an embodiment of the present invention also discloses a device for correcting fluorescence signal information of blood samples, please refer to FIG. 5 , the device for correcting fluorescence signal information of blood samples may include a memory 210 and The processor 230 and the memory 210 are used to store programs, and the processor 230 is used to execute the program stored in the memory 210 to implement the analysis method of any of the above-mentioned embodiments, for example to realize the above-mentioned steps 300, 330 and 350, and can also be used To implement step 370. The function of the processor 230 may be similar or the same as that of the processor 104 in the hematology analyzer. Specific instructions are given below.

In one embodiment of the processor 230, it acquires the fluorescent signal information of the blood sample treated with the fluorescent reagent, acquires the measured red blood cell parameters in the blood sample, and corrects the fluorescent signal information according to the red blood cell parameters . Fluorescence signals mainly represent white blood cell particles or ghost particles, where ghost particles refer to the particles of the plasma membrane that maintain the original shape and size of the red blood cells after the hypotonic treatment. In one embodiment, the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration.

As mentioned above, the data acquired by the processor 230 includes the fluorescence signal number information of the blood sample and the red blood cell parameters in the blood sample. In an embodiment, the processor 230 can also acquire the forward scattered light signal information or Side scattered light signal information. These three types of data—that is, the first type of data: the fluorescence information number information of the blood sample, the second type of data: the forward scattered light signal information or the side scattered light signal information of the blood sample, and the third type of data: all One or more of the red blood cell parameters in the blood sample—may be communicated to processor 230 by other devices.

The processor 230 correcting the fluorescent signal information according to the red blood cell parameters may include: performing an increase correction and/or a decrease correction on the fluorescent signal information according to the red blood cell parameters. The goal of the correction is: no matter how the red blood cell parameters such as HCT, RBC, and HGB change, the fluorescence signal value corresponding to the particle is corrected to be close to the normal fluorescence signal intensity value of the particle.

(1) Increase correction

The processor 230 performing increasing correction on the fluorescent signal information according to the red blood cell parameter includes: performing increasing correction on the fluorescent signal information if the red blood cell parameter is greater than a first preset value. Further, in one 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 magnitude of the increase correction performed by the processor 230 on the fluorescent signal information is. The setting of the first preset value can be based on the parameter value of normal red blood cells under the condition of normal fluorescence intensity of the particles as a reference. As mentioned above, the red blood cell parameters may include at least one of RBC, HGB and HCT. When the red blood cell parameters only include one of RBC, HGB and HCT, it is well understood that the red blood cell parameters are greater than the first preset value; when the red blood cell parameters include two or three of RBC, HGB and HCT, the red blood cell parameters are greater than The first preset value may refer to: performing a weighted sum of two or three of RBC, HGB, and HCT included in the red blood cell parameters, or a weighted average calculation. The specific weight value can be flexibly set in practice, and the calculated The value of is used as the red blood cell parameter to compare with the first preset value, and when the calculated value is greater than the first preset value, it is considered that the red blood cell parameter is greater than the first preset value. For example, when the red blood cell parameters include RBC, HGB and HCT, the weighted sum of the three parameters of RBC, HGB and HCT is calculated, or the weighted average is calculated, and the calculated value is regarded as the value of the red blood cell parameter to be compared with the first The preset value is compared, and when the calculated value is greater than the first preset value, it means that the red blood cell parameter is greater than the first preset value.

(2) Reduce correction

The processor 230 performing reduction correction on the fluorescent signal information according to the red blood cell parameter includes: performing reduction correction on the fluorescent signal information if the red blood cell parameter is smaller than a first preset value. Further, in one embodiment, if the red blood cell parameter is smaller than the first preset value, the smaller the red blood cell parameter is, the larger the reduction and correction of the fluorescent signal information by the processor 230 will be.

As mentioned above, the red blood cell parameters may include at least one of RBC, HGB and HCT. When the red blood cell parameters include only one of RBC, HGB and HCT, it is easy to understand that the red blood cell parameters are less than the first preset value; when the red blood cell parameters include two or three of RBC, HGB and HCT, the red blood cell parameters are less than The first preset value may refer to: performing a weighted sum of two or three of RBC, HGB, and HCT included in the red blood cell parameters, or a weighted average calculation. The specific weight value can be flexibly set in practice, and the calculated The value of is used as the red blood cell parameter to compare with the first preset value, and when the calculated value is smaller than the first preset value, it is considered that the red blood cell parameter is smaller than the first preset value. For example, when the red blood cell parameters include RBC, HGB and HCT, the weighted sum of the three parameters of RBC, HGB and HCT is calculated, or the weighted average is calculated, and the calculated value is regarded as the value of the red blood cell parameter to be compared with the first Compared with the preset value, when the calculated value is smaller than the first preset value, it means that the red blood cell parameter is smaller than the first preset value.

The processor 230 performs an increase correction and/or a decrease correction on the fluorescent signal information according to the red blood cell parameters, including: substituting the red blood cell parameters and the fluorescent signal information as independent variables into a preset correction function to obtain the correction The final fluorescent signal information, wherein the correction function is an increasing function. In an embodiment, the correction function may be a linear function or a power function. In one embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor uses the red blood cell parameters and the fluorescent signal information as an automatic Substituting variables into the preset correction function includes: substituting the red blood cell parameters as an independent variable into the first function to obtain a correction coefficient, using the correction coefficient as a constant and the fluorescent signal information as an independent variable into the second function In order to obtain the corrected fluorescent signal information. In an embodiment, both the first function and the second function are linear functions. For examples of the correction function, the first function and the second function, reference may be made to Embodiment 1, or specific embodiments which will be described in detail below, and will not be repeated here.

In an embodiment of the processor 230, it can also output the corrected fluorescence signal information, for example, display the fluorescence signal scatter diagram on the display according to the corrected fluorescence signal information, or send the corrected fluorescence signal information to the remote server , Central Station.

Of course, in some embodiments, after the correction, the processor 230 can also output the fluorescence signal information before correction and after correction, for example, displaying the corrected fluorescence signal scattergram on the display according to the corrected fluorescence signal information, and according to Fluorescent signal information before correction is displayed on the display as a scatter diagram of fluorescent signal before correction, which is convenient for users to compare.

In some embodiments, after correction, the processor 230 may also issue a reminder that the fluorescent signal information has been corrected, for example, send a reminder to the user through a display. In some embodiments, the processor 104 may also output the correction range of the fluorescence signal information, for example, display the correction range of the fluorescence signal information in a percentage form through a display. In some embodiments, after correction, if the correction magnitude of the fluorescent signal information exceeds a set threshold, a prompt or an alarm is sent out that the fluorescent signal information has been corrected.

The correction of fluorescent signal information by the present invention depends on red blood cell parameters, such as one or more of HCT, RBC and HGB. In the analysis of blood cells, red blood cell parameters such as HCT, RBC, and HGB are routinely measured parameters, so the correction of fluorescent signal information can be completed without increasing reagent consumption and measuring channels, so that the measurement cost will not Increase, and at the same time improve the accuracy of measurement results of measurement channels that require fluorescent signal information—such as NRBC channels, RET channels, and DIFF channels. For example, WBC counts, NRBC classification, Baso classification and other results can be given more accurately.

The following examples illustrate how the present invention corrects the fluorescent signal information according to the red blood cell parameters.

In the following, when explaining the relationship between the fluorescent signal information and the red blood cell parameters, it is mainly to study the relationship between the fluorescent signal of the WBC particle cluster and the red blood cell parameters in the experiment. The applicant counted 482 samples, took the HCT, RBC, and HGB of these samples as the horizontal axis, and took the position of the WBC particle cluster in the fluorescence direction as the vertical axis to draw a scatter diagram, as shown in Figure 6, Figure 7 and Figure 8 shown.

From the scatter plots in Figure 6 to Figure 8, it can be seen that the position value of WBC particle clusters in the fluorescence direction is negatively correlated with HCT, RBC, and HGB, that is, the smaller the HCT, RBC, and HGB, the smaller the WBC particle clusters are in the fluorescence direction. The larger the position value in the direction; the larger the HCT, RBC, and HGB, the smaller the position value of the WBC particle group in the fluorescence direction.

One or more of HCT, RBC and HGB can be used to calculate a correction coefficient, and then use the calculated correction coefficient to correct the fluorescence signal information, for example multiply the correction coefficient by the fluorescence signal information to obtain the corrected Fluorescent signal information. Taking Fig. 6 to Fig. 8 as an example, the goal of the correction is to make the position value of the WBC cluster in the direction of fluorescence basically unchanged with the changes of parameters such as HCT, RBC, and HGB. In Figures 6 to 8, we can set the position value of the WBC particle cluster in the fluorescence direction equal to 2000 as the target horizontal line (that is, the normal fluorescence signal intensity value of the white blood cell cluster, and the value varies according to different systems), that is, no matter How do the parameters of HCT, RBC, and HGB change, and the position value of the WBC particle group in the direction of fluorescence is basically maintained around 2000. With the change of HCT, RBC, 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 Figure 6, the first preset value corresponding to HCT is around 29; in Figure 7, the first preset value corresponding to RBC is around 2.3; in Figure 8, the first preset value corresponding to HGB is around 85. The setting of the first preset value can be based on the normal erythrocyte parameter value under the condition of normal fluorescence intensity of the particle as a reference, which can be understood as the corresponding erythrocyte parameter value when the correction coefficient is 1.

Let’s take HCT as an example to correct the fluorescent signal information in NRBC. According to the fitting results in Figure 6 above, the applicant constructed the first function f ₁₁ (x ₁ )=0.038x ₁ +0.09, where x ₁ represents the HCT value, the correction coefficient can be calculated by substituting the value of HCT; as shown in Figure 9, the 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 the HCT, the larger the correction coefficient The larger it is, the first function is an increasing function. After calculating the correction coefficient, multiply the fluorescence information signal by the correction coefficient to obtain the corrected fluorescence signal information, for example, the second function f ₁₂ (x _fl )=k ₁ x _fl +b, where k ₁ =f ₁₁ (x ₁ )=0.038x ₁ +0.09, x _fl represents the fluorescent signal information before correction; therefore the correction function is:

y _fl =f ₁ (x ₁ ,x _fl )=(0.038x ₁ +0.09)*x _fl +b;

Wherein y _fl represents the corrected fluorescent signal information; b is a constant, for example, 0; the symbol * is a multiplication sign.

The first function used for correction may correspond to a linear function, or may correspond to other functional forms, such as a power function. In short, the correction target is no matter how the parameters of HCT, RBC, and HGB change, the position value of the WBC particle cluster in the fluorescence direction is basically maintained near the target line, and the fluorescence signal value corresponding to the particle is corrected to the normal fluorescence signal intensity of the particle. near the value.

Figure 10 shows the scatter diagram of the cell particles of the three samples in the NRBC channel (mainly the WBC particle cluster in the upper right corner and the ghost particle cluster in the lower left corner), where the abscissa is the value of the fluorescence signal information of the cell particles , and the ordinate is the forward scatter light signal information of the cell particle. Next, according to the above-mentioned correction function y _fl =f ₁ (x ₁ ,x _fl )=(0.038x ₁ +0.09)*x _fl +b, the fluorescence signal information of sample 1, sample 2 and sample 3 is respectively corrected, where b Take it as 0.

In sample 1, RBC=0.76×10.^12/L, HGB=26g/L, HCT=8.2%. Figure 10(a) is the scatter diagram of the uncorrected cell particles obtained in the NRBC channel. It can be seen that the fluorescence signal information of the cell particle cluster is too large; the HCT value of sample 1 is 8.2%, which is brought into the first After the function f ₁₁ (x ₁ )=0.038x ₁ +0.09, the correction coefficient is 0.4016. By multiplying the correction coefficient 0.4016 by the value of the fluorescence signal information of each cell particle in sample 1, the corrected value of the fluorescence signal information of each cell particle can be obtained, as shown in Figure 10(d).

In sample 2, RBC=3.25×10.^12/L, HGB=71g/L, HCT=23.8%. Figure 10(b) is the scatter diagram of the uncorrected cell particles obtained in the NRBC channel. It can be seen that the fluorescence signal information of the WBC particle cluster is moderate; the HCT value of sample 2 is 23.8%, which is brought into the first function After f ₁₁ (x ₁ )=0.038x ₁ +0.09, the correction coefficient is 0.9944. By multiplying the correction coefficient 0.9944 by the value of the fluorescence signal information of each cell particle in sample 2, the corrected value of the fluorescence signal information of each cell particle can be obtained, as shown in Figure 10(e).

In sample 3, RBC=7.04×10.^12/L, HGB=158g/L, HCT=50.3%. Figure 10(c) is the scatter diagram of the uncorrected cell particles obtained in the NRBC channel. It can be seen that the fluorescence signal information of the cell particle cluster is relatively small; the HCT value of sample 3 is 50.3%, which is brought into the first After the function f ₁₁ (x ₁ )=0.038x ₁ +0.09, the correction factor is 2.0014. By multiplying the correction coefficient 2.0014 by the value of the fluorescence signal information of each cell particle in sample 3, the corrected value of the fluorescence signal information of each cell particle can be obtained, as shown in FIG. 10(f).

From Figures 10(e) to 10(f), it can be seen that the fluorescent signals of the cell particle clusters are all corrected to a more appropriate range, that is, a relatively central position in the scatter diagram.

Due to the dye competition of NRBC channel, BASO channel, RET channel, DIFF channel and other channel dye methods, the fluorescent signal of white blood cell population is abnormal, which may easily lead to errors in the classification of white blood cell population or NRBC population. In the classification of white blood cells, the applicant has discovered through long-term research and development that, in some samples, when the cells are treated with fluorescent dyes and the cells are classified using fluorescent signals and scattered light signals, there will be deviations between the measurement results and the results of microscopic examination . Through in-depth research, it was found that in such samples, the parameters of red blood cells were different from normal values. Through verification, it is found that there is a certain correlation between the red blood cell parameters and the intensity of the fluorescent signal, and the red blood cell parameters can be used to correct the fluorescent signal. While not being bound by theory, the applicant speculates that red blood cells may affect the staining behavior of other cells that need to be sorted, resulting in abnormal fluorescent signals.

The embodiment of the present application corrects the fluorescent signal according to the red blood cell parameters, thereby correcting the position of the white blood cell group in the scatter diagram, and solving the problem of inaccurate classification of the white blood cell group. In white blood cell classification, taking the fixed demarcation algorithm as an example, the embodiment of the present application can allow the target particle group, such as white blood cell group, or nucleated red blood cell group, or reticulocyte group, to be accurately located in the predetermined area of the fixed demarcation algorithm. In the designated area, the accuracy of classification and counting is guaranteed. This application can correct the fluorescent signal with red blood cell parameters under the NRBC channel, BASO channel, WNB channel, RET channel, and DIFF channel, so as to correct the position of the white blood cell group in the scatter diagram.

Those skilled in the art can understand that all or part of the functions of the various methods in the foregoing implementation manners can be realized by means of hardware, or by means of computer programs. When all or part of the functions in the above embodiments are implemented by means of a computer program, the program can be stored in a computer-readable storage medium, and the storage medium can include: read-only memory, random access memory, magnetic disk, optical disk, hard disk, etc., through The computer executes the program to realize the above-mentioned functions. For example, the program is stored in the memory of the device, and when the processor executes the program in the memory, all or part of the above-mentioned functions can be realized. In addition, when all or part of the functions in the above embodiments are realized by means of a computer program, the program can also be stored in a storage medium such as a server, another computer, a magnetic disk, an optical disk, a flash disk, or a mobile hard disk, and saved by downloading or copying. To the memory of the local device, or to update the version of the system of the local device, when the processor executes the program in the memory, all or part of the functions in the above embodiments can be realized.

The above uses specific examples to illustrate the present invention, which is only used to help understand the present invention, and is not intended to limit the present invention. For those skilled in the technical field to which the present invention belongs, some simple deduction, deformation or replacement can also be made according to the idea of the present invention.

Claims (29)

1. An analysis method for a blood sample, comprising: Obtain the fluorescence signal information of the blood sample treated with the fluorescent reagent; Acquire the measured red blood cell parameters in the blood sample; the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration; The fluorescent signal information is corrected according to the red blood cell parameters and/or an abnormality prompt is issued for the fluorescent signal information. 2. The analysis method of a blood sample according to claim 1, wherein said acquiring the fluorescent signal information of the blood sample treated with the fluorescent reagent comprises: acquiring the fluorescent signal information of the blood sample treated with the fluorescent reagent, And forward scattered light signal information and/or side scattered light signal information. 3. The analysis method of a blood sample according to claim 2, wherein said obtaining the fluorescent signal information of the blood sample processed by the fluorescent reagent comprises: measuring the blood sample by flow cytometry to obtain Fluorescent signal information of the blood sample treated with the fluorescent reagent, forward scattered light signal information and/or side scattered light signal information. 4. The blood sample analysis method according to claim 2 or 3, further comprising: performing leukocyte classification and/or counting according to the side scattered light value of the blood sample and the corrected fluorescent signal information. 5. The blood sample analysis method according to claim 2 or 3, further comprising: counting nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescent signal information. 6. The blood sample analysis method according to claim 2 or 3, further comprising: performing leukocyte count and/or reticulation according to the forward scattered light value of the blood sample and the corrected fluorescent signal information Red blood cell identification and/or basophil classification. 7. The blood sample analysis method according to claim 2 or 3, further comprising: performing basophil classification according to the side scattered light value of the blood sample and the corrected fluorescence signal information. 8. The analysis method of blood samples according to claim 1, characterized in that, before obtaining the fluorescent signal information of the blood samples processed by the fluorescent reagents, further comprising: measuring red blood cell parameters on the blood samples to determine the red blood cell parameters, and then perform hemolysis treatment and fluorescent reagent treatment on the blood sample, and measure the fluorescent signal information of the blood sample. 9. The analysis method of a blood sample according to claim 8, wherein said measuring the red blood cell parameters of the blood sample to determine the red blood cell parameters comprises: using a small hole impedance method or an optical method to measure the red blood cell parameters. A blood sample is subjected to red blood cell parameter measurements to determine said red blood cell parameter. 10. The method for analyzing blood samples according to claim 1, wherein said correcting said fluorescent signal information according to said red blood cell parameters comprises: using said red blood cell parameters and said fluorescent signal information as independent variables Substitute the preset correction function to obtain the corrected fluorescent signal information as the dependent variable. 11. The blood sample analysis method according to claim 10, wherein the correction function is an increasing function of the dependent variable relative to the independent variable. 12. The blood sample analysis method according to claim 10, wherein the correction function is a linear function, a polynomial function or a power function. 13 . The blood sample analysis method according to claim 1 , further comprising: outputting the corrected fluorescent signal information after correcting the fluorescent signal information according to the red blood cell parameters. 14 . 14 . The blood sample analysis method according to claim 13 , further comprising: outputting the fluorescent signal information before and after correction after correcting the fluorescent signal information according to the red blood cell parameters. 15 . 15. The blood sample analysis method according to claim 1, characterized in that after correcting the fluorescent signal information according to the red blood cell parameters, further comprising: issuing a reminder that the fluorescent signal information has been corrected or issuing an alarm. 16 . The blood sample analysis method according to claim 15 , wherein the prompting that the fluorescent signal information has been corrected comprises: outputting a correction range of the fluorescent signal information. 17 . 17. The blood sample analysis method according to claim 15, characterized in that, after correcting the fluorescent signal information according to the red blood cell parameters, further comprising: if the correction range of the fluorescent signal information exceeds a set threshold, Then send out the reminder that the fluorescent signal information has been corrected or send out an alarm. 18. A blood cell analyzer, comprising: At least one reaction pool is used to provide a reaction place for blood samples and reagents; The optical detection device is used to irradiate the blood sample treated with the reagent with light, collect the optical signal generated by each particle in the blood sample treated with the reagent due to the light irradiation, and convert it into an electrical signal to output the optical signal information; A delivery device, used to deliver the reagent-treated blood sample in the reaction cell to the optical detection device; a processor, configured to receive and process the optical signal information output by the optical detection device to obtain the measurement parameters of the blood sample; According to the red blood cell parameters in the blood sample, the fluorescent signal information is corrected according to the red blood cell parameters and/or abnormal fluorescent signal information is issued; the red blood cell parameters include at least one of hematocrit, red blood cell count and hemoglobin concentration kind. 19. The blood cell analyzer according to claim 18, wherein the processor acquires the fluorescent signal information of the blood sample treated with the fluorescent reagent, and the forward scattered light signal information and/or side to the scattered light signal information. 20. The blood cell analyzer according to claim 19, wherein the optical detection device comprises a light source and a flow chamber having an orifice, and the leukocyte particles in the blood sample treated with the fluorescent reagent are in the flow in the flow chamber and pass through the orifice one by one, the light emitted by the light source irradiates the particles in the orifice and correspondingly generates fluorescent signals and scattered light signals, which are converted by the optical detection device to the processing The detector outputs fluorescence signal information, and forward scattered light signal information and/or side scattered light signal information. 21. The blood cell analyzer according to claim 19 or 20, wherein the processor further classifies and/or counts white blood cells according to the side scattered light value of the blood sample and the corrected fluorescent signal information . 22. The blood cell analyzer according to claim 19 or 20, wherein the processor also counts nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescent signal information. 23. The blood cell analyzer according to claim 19 or 20, wherein the processor also performs white blood cell counting and/or network analysis according to the forward scattered light value of the blood sample and the corrected fluorescent signal information. Erythrocyte identification and/or basophil classification. 24. The blood cell analyzer according to claim 19 or 20, wherein the processor further classifies basophils according to the side scattered light value of the blood sample and the corrected fluorescent signal information . 25. The blood cell analyzer according to claim 18, characterized in that, the reaction pool provides a reaction place for the blood sample and the reagents required for red blood cell parameter measurement, and the optical detection device reacts to the reagents processed by the reagents After the red blood sample is measured, and the optical signal information is output, the processor determines the red blood cell parameter according to the optical signal information; the reaction pool performs hemolysis treatment and fluorescent reagent treatment on the blood sample, and then the The optical detection device measures the processed blood sample, and outputs the fluorescence signal information of the blood sample. 26. The blood cell analyzer according to claim 18, wherein the processor is configured to: substitute the red blood cell parameters and the fluorescent signal information as independent variables into a preset correction function to obtain The corrected fluorescent signal information. 27. The blood cell analyzer according to claim 26, wherein the correction function is an increasing function of the dependent variable relative to the independent variable. 28. The blood cell analyzer according to claim 18, wherein the processor also outputs the corrected fluorescent signal information. 29. A computer-readable storage medium, wherein a program is stored, and the program can be executed by a processor to implement the method according to any one of claims 1-17.

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