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

Analysis method of blood sample, blood cell analyzer and storage medium Download PDF

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CN110887818B
CN110887818B CN201811045480.8A CN201811045480A CN110887818B CN 110887818 B CN110887818 B CN 110887818B CN 201811045480 A CN201811045480 A CN 201811045480A CN 110887818 B CN110887818 B CN 110887818B
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signal information
blood sample
blood cell
red blood
fluorescent
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CN110887818A (en
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祁欢
王官振
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Shenzhen Mindray Bio Medical Electronics Co Ltd
Shenzhen Mindray Scientific Co Ltd
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Shenzhen Mindray Bio Medical Electronics Co Ltd
Shenzhen Mindray Scientific Co Ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N2015/1486—Counting the particles

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

Analysis method of blood sample, 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
A blood cell analyzer is an instrument that can detect cells in blood, for example, can classify and count white blood cells, red blood cells, platelets, nucleated red blood cells, reticulocytes, and the like in blood.
Taking white blood cell detection as an example, one of the most common methods is laser light scattering, i.e., a blood cell analyzer uses scattered light to sort and count white blood cells. The scattered light includes three optical signals, forward scattered light, side scattered light, and fluorescent signal. In general, forward scattered light may reflect cell size information, side scattered light may reflect the complexity of the internal structure of the cell, and fluorescent signals may reflect the content of substances such as DNA, RNA, etc. within the cell that may be stained by fluorescent dyes. 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 white blood cell classification counting method is used for some blood tests, the accuracy can not meet the requirements. Therefore, development of a white blood cell classification method with better accuracy is required.
Disclosure of Invention
The application provides a blood sample analysis method, a blood cell analyzer and a storage medium.
There is provided a method of analyzing a blood sample, comprising:
obtaining fluorescent 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.
In one embodiment, the obtaining fluorescence signal information of the blood sample after the fluorescent reagent treatment comprises: and acquiring fluorescence signal information, forward scattering light signal information and/or side scattering light signal information of the blood sample treated by the fluorescent reagent.
In one embodiment, the obtaining fluorescence signal information of the blood sample after the fluorescent reagent treatment comprises: and measuring the blood sample by adopting flow cytometry to obtain fluorescence signal information, forward scattering light signal information and/or side scattering light signal information of the blood sample treated by the fluorescent reagent.
In one embodiment, the method further comprises: and performing 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: and counting 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: and performing white blood cell count and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value and the corrected fluorescence signal information of the blood sample.
In one embodiment, the method further comprises: and classifying basophils according to the side scattering light value of the blood sample and the corrected fluorescence signal information.
In one embodiment, before obtaining the fluorescence signal information of the blood sample treated by the fluorescent reagent, the method further comprises: and performing red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, then performing hemolysis treatment and fluorescent reagent treatment on the blood sample, and measuring fluorescent signal information of the blood sample.
In one embodiment, the performing red blood cell parameter measurements on the blood sample to determine the red blood cell parameter comprises: and measuring the red blood cell parameters of the blood sample by adopting a small hole impedance method or an optical method to determine the red blood cell parameters.
In one embodiment, said modifying said fluorescence signal information according to said red blood cell parameter comprises: substituting the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as the 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 correction function is a linear function, a polynomial function, or a power function.
In one embodiment, the red blood cell parameter comprises at least one of hematocrit, red blood cell count, and hemoglobin concentration.
In one embodiment, after correcting the fluorescence signal information according to the red blood cell parameter, the method further comprises: outputting the corrected fluorescence signal information.
In one embodiment, after correcting the fluorescence signal information according to the red blood cell parameter, the method further comprises: outputting the fluorescence signal information before and after correction.
In one embodiment, 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.
In one embodiment, the prompting that the fluorescence signal information is corrected comprises: and outputting the corrected amplitude of the fluorescence signal information.
In one embodiment, after correcting the fluorescence signal information according to the red blood cell parameter, the method further comprises: and if the corrected amplitude of the fluorescent signal information exceeds the set threshold value, sending a prompt that the fluorescent signal information is corrected or sending an alarm.
There is also provided a blood cell analyzer comprising:
at least one reaction cell for providing a reaction site for a blood sample and a reagent;
the optical detection device is used for carrying out light irradiation on the blood sample after the reagent treatment, collecting optical signals generated by each particle in the blood sample after the reagent treatment 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 tank to the optical detection device;
the processor is used for receiving and processing the optical signal information output by the optical detection device so as to obtain the measurement parameters of the blood sample; the processor acquires fluorescent signal information of the blood sample treated by the fluorescent reagent, acquires the measured red blood cell parameters in the blood sample, corrects the fluorescent signal information according to the red blood cell parameters and/or sends out abnormal prompt of the fluorescent signal information.
In one embodiment, the processor obtains fluorescence signal information of the blood sample after treatment with the fluorescent reagent, and forward scattered light signal information and/or side scattered light signal information of the leukocyte particles.
In one embodiment, the optical detection device comprises a light source and a flow chamber with an orifice, white blood cell particles in the blood sample treated by the fluorescent reagent flow in the flow chamber and pass through the orifice one by one, the particles in the orifice are irradiated by light emitted by the light source and correspondingly generate fluorescent signals and scattered light signals, and fluorescent signal information and forward scattered light signal information and/or side scattered light signal information are output to the processor after being converted by the optical detection device.
In one embodiment, the processor further performs white blood cell classification and/or counting based on the side scatter light values and the modified fluorescent signal information of the blood sample.
In one embodiment, the processor further performs nucleated red blood cell counting based on the forward scattered light values of the blood sample and the corrected fluorescence signal information.
In one embodiment, the processor further performs white blood cell count and/or reticulocyte identification and/or basophil classification based on the forward scattered light value and the modified fluorescent signal information of the blood sample.
In one embodiment, the processor further classifies basophils based on the side scatter light values and modified fluorescent signal information of the blood sample.
In one embodiment, the reaction tank provides a reaction place for the blood sample and a reagent required by the blood sample for measuring the erythrocyte parameters, the optical detection device measures the erythrocyte parameters of the blood sample treated by the reagent and outputs optical signal information, and the processor determines the erythrocyte parameters according to the optical signal information; and the reaction tank carries out hemolysis treatment and fluorescent reagent treatment on the blood sample, and then the optical detection device measures the treated blood sample and outputs fluorescent signal information of the blood sample.
In one embodiment, the processor is configured to: substituting the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as the 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 red blood cell parameter comprises at least one of hematocrit, red blood cell count, and hemoglobin concentration.
In one embodiment, the processor further outputs modified fluorescent 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 analysis method of the blood sample, the blood cell analyzer and the computer readable storage medium of the embodiment, the red blood cell parameter is used for correcting the fluorescence signal information, so that the accuracy of the fluorescence signal information is improved.
Drawings
FIG. 1 is a schematic diagram 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 diagram showing a structure of a fluorescence signal information modifying apparatus for a blood sample according to an embodiment;
FIG. 6 is a scatter plot of HCT versus position values of WBC clusters in the fluorescent direction for a plurality of samples;
FIG. 7 is a scatter plot of the position values of WBC clusters in the fluorescent direction versus RBC for a plurality of samples;
FIG. 8 is a scatter plot of the position values of WBC clusters in the fluorescent direction versus HGB for a plurality of samples;
FIG. 9 is a functional diagram of a first function modified using HCT, in accordance with one embodiment;
FIG. 10 is a schematic representation of fluorescence signal information of pre-and post-correction cellular particles in three samples.
Detailed Description
The invention will be described in further detail below with reference to the drawings by means of specific embodiments. Wherein like elements in different embodiments are numbered alike in association. In the following embodiments, numerous specific details are set forth in order to provide a better understanding of the present application. However, one skilled in the art will readily recognize that some of the features may be omitted, or replaced by other elements, materials, or methods in different situations. In some instances, some operations associated with the present application have not been shown or described in the specification to avoid obscuring the core portions of the present application, and may not be necessary for a person skilled in the art to describe in detail the relevant operations based on the description herein and the general knowledge of one skilled in the art.
Furthermore, the described features, operations, or characteristics of the description may be combined in any suitable manner in various embodiments. Also, various steps or acts in the method descriptions may be interchanged or modified in a manner apparent to those of ordinary skill in the art. Thus, the various orders in the description and drawings are for clarity of description of only certain embodiments, and are not meant to be required orders unless otherwise indicated.
The numbering of the components itself, e.g. "first", "second", etc., is used herein merely to distinguish between the described objects and does not have any sequential or technical meaning. The terms "coupled" and "connected," as used herein, are intended to encompass both direct and indirect coupling (coupling), unless otherwise indicated.
Taking a common five-class blood cell analyzer as an example, the device generally has a channel requiring fluorescent signal measurement, and uses three scattered light signals such as forward scattered light, side scattered light, fluorescent signal and the like to conduct cell classification and counting, for example, white blood cell counting, nucleated red blood cell classification, and some basophil classification can be conducted.
Some english terms of the present application will be described first.
RBC: red blood cells (number);
NRBC: nucleated red blood cells (number);
RET: reticulocytes (number);
HGB: erythrocyte hemoglobin content;
HCT: hematocrit;
MCV: average red blood cell volume;
WBC: white blood cells (number);
BASO: basophils (numbers).
In a common five-class blood cell analyzer, a channel requiring fluorescent signal measurement exists, and three optical signals such as forward scattered light, side fluorescent signal and the like are adopted to carry out cell classification and counting, for example, white blood cell counting, nucleated red blood cell classification and basophil classification can be carried out. Herein, a channel refers to a detection channel for identifying and/or counting and/or classifying a certain type of cells, for example, an NRBC channel is a channel for counting nucleated red blood cells, a WNB channel is a channel for classifying nucleated red blood cells, a RET channel is a channel for classifying and/or counting reticulocytes, a DIFF channel is a channel for classifying and/or counting white blood cells, and a BASO channel is a channel for counting basophils, which are known to those skilled in the art and are not described in detail herein.
Classification and counting by acquiring optical signals of cells is erroneous when WBC clusters of different samples are in different positions. In the classification of white blood cells, the applicant has found through long-term research and development that under some samples, when cells are treated with a fluorescent dye, the measurement result deviates from the microscopic examination result when the cells are classified by using a fluorescent signal and a scattered light signal. Through intensive studies, it was found that parameters of erythrocytes in such samples are different from normal values. Through verification, the red blood cell parameter has a certain correlation with the fluorescence signal intensity, and the fluorescence signal can be corrected by using the red blood cell parameter. While not being bound by theory, the applicant speculates that it is possible that erythrocytes will affect the staining behavior of other cells to be classified, resulting in abnormal fluorescent signals.
In practice, the applicant found that, in the current five-class blood cell analyzer, the NRBC channel is taken as an example, when the cells are stained by using fluorescent dye, the positions of the fluorescent signals of the cells of different samples in the fluorescent direction are different, and in the process of counting and classifying the cells (such as white blood cells), the counting and classifying gating white blood cell algorithm is influenced (such as the fluorescent signal influencing gating), which influences the counting and classifying of the NRBC channel, and further leads to the increase of the risk of clinical misdiagnosis. Taking RET channel as an example, the treatment degree of fluorescent dye on cells is different due to red blood cells with different distributions in the testing process, so that signals of cells in fluorescence direction are different, and influence on classifying red blood cells such as RET identification is caused.
Therefore, for those channels using fluorescent dyes, such as NRBC channels, RET channels, DIFF channels, etc., when these channels perform 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 a limited number of bytes of the signal collector, because the signal will have a maximum and minimum limit, the situation that WBC clusters are too far left or right in the scattergram will eventually occur, thus resulting in distortion of the scattergram of the channel, missing necessary scattergram information, causing difficulty for subsequent classification, and classification errors of ghosts, nucleated erythrocytes, basophils will easily occur, thus giving erroneous counting and classification results.
In view of the above, there is a need for a solution for correcting the difference in fluorescence signals of different samples in channels employing fluorescent dyes, such as NRBC channel, BASO channel, WNB channel, RET channel, and DIFF channel.
The applicant is directed to the above problems, and after the theoretical analysis of reagent action and the research of a large number of samples, the invention provides that for the channel using fluorescent dye, such as NRBC channel, in the actual measurement process, the fluorescence signal of WBC particles is in negative correlation with red blood cell count (RBC), red blood cell hemoglobin content (HGB) and red blood cell packed volume (HCT), i.e. the smaller the same sample or blood sample, the larger the fluorescence signal of WBC particles; the larger the RBC, HGB, HCT, the smaller the WBC particle fluorescence signal. Therefore, RBC, HGB, HCT can be utilized to correct the fluorescence signal of the WBC particle and/or send out abnormal indication of the fluorescence signal information, namely, the fluorescence signal information can be corrected according to the red blood cell parameter and/or send out abnormal indication of the fluorescence signal information. The correction may be performed only, or only the abnormality notification may be issued, or both the correction and the abnormality notification may be performed.
Example 1
The blood cell analyzer according to an embodiment of the present invention may mainly include the 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, described in detail below.
The reaction cell 101 is used to provide a reaction site for a blood sample and a reagent to prepare a sample liquid. Specifically, a blood sample obtained by blood sampling may be diluted and labeled with a fluorescent staining reagent to obtain a sample liquid. 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 sample solution, which is the blood sample treated with the reagent, collect optical signals generated by the irradiation of each particle in the blood sample treated with the reagent, and convert the optical signals into electrical signals to output optical signal information. The optical signal may be a forward scattered light signal, a side scattered light signal, a fluorescent signal. In one embodiment of the optical detection device 102, which may include a light source 1021 and a sheath flow chamber 1022 having an aperture 10221, particles in the blood sample may flow within the sheath flow chamber 1022 and pass through the aperture 10221 one by one, light emitted by the light source 1021 may impinge on the particles in the aperture 10221 and generate a scattered light signal and/or a fluorescent signal, respectively. The optical detection device 102 may further include a lens group 1023 disposed in front of and laterally of the aperture, a photo-sensor 1024 (e.g., photodiode, photomultiplier tube, etc.), and an a/D converter, which may be disposed in the processor 104 or separately formed as a component, so that the lens group 1023 captures the corresponding scattered light signal and fluorescent signal, the photo-sensor 1024 converts the captured optical signal (e.g., scattered light signal and fluorescent signal) into an electrical signal, and the a/D converter processes the electrical signal into a digital signal via a/D conversion, and outputs the digital signal as optical signal information.
The transfer device 103 is used to transfer the sample liquid, which is the blood sample treated with the reagent in the reaction cell 101, 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 a measured parameter of the blood sample. In one embodiment of the processor 104, it obtains fluorescent signal information of the blood sample after being treated by the fluorescent reagent, obtains measured red blood cell parameters in the blood sample, and corrects the fluorescent signal information according to the red blood cell parameters. The particles represented by the fluorescent signals are mainly white blood cell particles or ghost particles, wherein the ghost particles are particles of which the plasma membrane is broken to remain the original form and size of the cell membrane structure after the red blood cells are subjected to hypotonic treatment. In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.
As described above, the data acquired by the processor 104 includes fluorescence information of the blood sample and red blood cell parameters in the blood sample, and in an embodiment, the processor 104 may also acquire forward scattered light signal information or side scattered light signal information of the blood sample. These three types of data-namely, the first type of data: fluorescence information number information of the blood sample, second kind of data: forward scattered light signal information or side scattered light signal information of the blood sample, data of a third type: one or more of the red blood cell parameters in the blood sample may be transmitted by other devices to the processor in the blood cell analyzer of the present invention or may be measured by the blood cell analyzer of the present invention, as exemplified below.
Processor 104 obtains a first type of data (fluorescence information number information of the blood sample): the reaction cell 101 performs hemolysis and fluorescent reagent treatment on a blood sample, the conveying device 103 conveys the blood sample subjected to the hemolysis and fluorescent reagent treatment to the optical detection device 102, the optical detection device 102 measures the blood sample subjected to the hemolysis and fluorescent reagent treatment, and fluorescence signal information of the blood sample is output to the processor 104.
Processor 104 obtains second type data (forward scattered light signal information or side scattered light signal information of the blood sample): the forward scattered light signal information and the side scattered light signal information can be acquired when the fluorescence signal is acquired; for example, the reaction cell 101 performs a hemolysis treatment and a fluorescent reagent treatment on a blood sample, the transfer device 103 transfers the blood sample after the hemolysis treatment and the fluorescent reagent treatment to 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 one or both of fluorescence signal information, forward scattered light signal information and side scattered light signal information of the blood sample to the processor 104. Specifically, by the conveyance device 103, the cell particles in the blood sample after the fluorescent reagent treatment 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 a fluorescent signal and a scattered light signal (refer to one or both of forward scattered light signal information and side scattered light signal information) of the cell particles correspondingly, and the fluorescent signal information, and the forward scattered light signal information or the side scattered light signal information are output to the processor 104 after the conversion by the optical detection device 10221.
The processor 104 can perform the following cell detection by acquiring fluorescence signal information and scattered light signal information. White blood cell classification and/or enumeration is performed based on the side scatter light values and modified fluorescent signal information of the blood sample, e.g., white blood cell tetra classification via DIFF channels. The nucleated red blood cells are counted based on the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, through an NRBC channel. White blood cell count and/or reticulocyte identification and/or basophil classification are performed based on the forward scattered light value and the corrected fluorescence signal information of the blood sample, such as reticulocyte classification and/or counting via RET channels. Basophil classification is performed based on the side scatter light values and the corrected fluorescence signal information of the blood sample, for example, BASO channels.
Processor 104 obtains a third type of data (red blood cell parameters in the blood sample): the reaction cell 101 provides a reaction place for the blood sample and the reagent required by the measurement of the erythrocyte parameters, 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 parameters of the blood sample processed by the reagent, and outputs optical signal information, and the processor 104 determines the erythrocyte parameters according to the optical signal information. The red blood cell parameters in the blood sample can be detected and acquired by an optical method. Of course, in other embodiments, the red blood cell parameters in the blood sample may also be obtained by a small hole impedance measurement, such as from a detection device of the blood cell analyzer itself or other instrument.
The above description is given of the processor 104 acquiring the three types of data, and the following description is given of the correction 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 carrying out increasing correction and/or decreasing correction on the fluorescence signal information according to the red blood cell parameters. The objective of the correction is: regardless of how the HCT, RBC, HGB parameters of the red blood cells change, the fluorescence signal value corresponding to the particle is corrected to be near the normal fluorescence signal intensity value of the particle.
(1) Augmentation correction
The processor 104 performs augmentation correction on the fluorescence signal information according to the red blood cell parameter, including: and if the red blood cell 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 preset value, the greater the red blood cell parameter, the greater the magnitude by which the processor 104 performs the increase correction on the fluorescence signal information. The setting of the first preset value may be based on the normal red blood cell parameter value of the particle under normal fluorescence intensity.
As described above, the red blood cell parameter may comprise 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, a red blood cell parameter greater than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 greater than the first preset value, it means that the red blood cell parameter is greater than the first preset value.
(2) Reduction correction
The processor 104 performs 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 less than the first preset value, the smaller the red blood cell parameter, the greater the magnitude of the reduction correction performed by the processor 104 on the fluorescence signal information
As described above, the red blood cell parameter may comprise 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, a red blood cell parameter less than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 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 fluorescence signal information according to the red blood cell parameter, including: substituting the erythrocyte parameter 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 an embodiment, the correction function may be a linear function or a power function. In an embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor substitutes the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function, wherein the method comprises the following steps of: substituting the red blood cell parameter as an independent variable into the first function to obtain a correction coefficient, substituting the correction coefficient as a constant and substituting the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information as an independent variable. In one embodiment of the processor 104, the modified fluorescent signal information may also be output, for example, by displaying a fluorescent signal scatter plot via a display based on the modified fluorescent signal information, or by sending the modified fluorescent 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, the corrected fluorescence signal scatter diagram is displayed through the display according to the corrected fluorescence signal information, and the fluorescence signal scatter diagram before correction is displayed through the display according to the fluorescence signal information before correction, so as to facilitate comparison of users.
In some embodiments, after correction, the processor 104 may also issue a prompt or alert that the fluorescent signaling information has been corrected, such as by a display, to the user. In some embodiments, the processor 104 may also output the corrected amplitude of the fluorescence signal information, for example, by displaying the corrected amplitude of the fluorescence signal information in percentage form via a display. In some embodiments, a prompt or alarm is issued that the fluorescence signal information has been modified if the modified amplitude of the fluorescence signal information exceeds a set threshold.
The correction function, the first function, and the second function are exemplified below.
In the formulas of the following formulas (1) to (7), y fl Representing corrected fluorescence signal information, x fl Representing fluorescence signal information before correction, x 1 、x 2 、x 3 Each representing a value of HCT, RBC, HGB.
(1) When the erythrocyte parameter includes only HCT, the correction function may have the following form:
y fl ＝f 1 (x 1 ,x fl )；
further, the function f is modified 1 (x 1 ,x fl ) The first function included may be f 11 (x 1 ) The second function may be f 12 (x fl ) For example a second function f 12 (x fl )＝k 1 x fl +b，k 1 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 1 ＝f 11 (x 1 )。
(2) When the red blood cell parameter includes only RBCs, the correction function may have the form:
y fl ＝f 2 (x 2 ,x fl )；
Further, the function f is modified 2 (x 2 ,x fl ) The first function included may be f 21 (x 2 ) The second function may be f 22 (x fl ) For example a second function f 22 (x fl )＝k 2 x fl +b，k 2 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 2 ＝f 21 (x 2 )。
(3) When the erythrocyte parameter includes only HGB, the correction function may have the following form:
y fl ＝f 3 (x 3 ,x fl )；
further, the function f is modified 3 (x 3 ,x fl ) The first function included may be f 31 (x 3 ) The second function may be f 32 (x fl ) For example a second function f 32 (x fl )＝k 3 x fl +b，k 3 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 3 ＝f 31 (x 3 )。
(4) When the red blood cell parameters include HCT and RBC, the correction function may have the form:
y fl ＝f 4 (x 1 ,x 2 ,x fl )；
further, the function f is modified 4 (x 1 ,x 2 ,x fl ) The first function included may be f 41 (x 1 ,x 2 ) The second function may be f 42 (x fl ) For example a second function f 42 (x fl )＝k 4 x fl +b，k 4 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 4 ＝f 41 (x 1 ,x 2 )。
(5) When the red blood cell parameters include RBC and HGB, the correction function may have the following form:
y fl ＝f 5 (x 2 ,x 3 ,x fl )；
further, the function f is modified 5 (x 2 ,x 3 ,x fl ) The first function included may be f 51 (x 2 ,x 3 ) The second function may be f 52 (x fl ) For example a second function f 52 (x fl )＝k 5 x fl +b，k 5 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 5 ＝f 51 (x 2 ,x 3 )。
(6) When the erythrocyte parameters include HCT and HGB, the correction function may have the following form:
y fl ＝f 6 (x 1 ,x 3 ,x fl )；
further, the function f is modified 6 (x 1 ,x 3 ,x fl ) The first function included may be f 61 (x 1 ,x 3 ) The second function may be f 62 (x fl ) For example a second function f 62 (x fl )＝k 6 x fl +b，k 6 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 6 ＝f 61 (x 1 ,x 3 )。
(7) When the red blood cell parameters include HCT, RBC, and HGB, the correction function may have the form:
y fl ＝f 7 (x 1 ,x 2 ,x 3 ,x fl )；
further, the function f is modified 7 (x 1 ,x 2 ,x 3 ,x fl ) The first function included may be f 71 (x 1 ,x 2 ,x 3 ) The second function may be f 72 (x fl ) For example a second function f 72 (x fl )＝k 7 x fl +b，k 7 And b is a constant, and the above-mentioned substitution of the correction coefficient as a constant into the second function may be referred to as k 7 ＝f 71 (x 1 ,x 2 ,x 3 )。
Example 2
An embodiment of the present invention further provides an analysis method (hereinafter referred to as an analysis method) for a blood sample. Referring to FIG. 2, the analysis method may include steps 310-350.
Step 310: and obtaining fluorescence signal information of the blood sample treated by the fluorescent reagent. In one embodiment, step 310 may include: and acquiring fluorescence signal information and forward scattering light signal information or side scattering light signal information of the blood sample treated by the fluorescent reagent.
Step 330: obtaining measured parameters 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 scattered light signal information, and the side scattered 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 may be 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 may be obtained by 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.
Taking the example of measuring the fluorescence signal information of the obtained blood sample and the red blood cell parameters in the blood sample in real time, referring to fig. 3, one embodiment of the analysis method may further include a step 300 and a step 320, which are described in detail below.
Step 300: and performing hemolysis treatment and fluorescent reagent treatment on the blood sample, and measuring fluorescent signal information of the blood sample.
Step 320: red blood cell parameter measurements are performed on the blood sample to determine the red blood cell parameter. In an embodiment, if the analysis method has both step 300 and step 320, it may be that step 320 is performed first, and then step 300 is performed, for example, performing red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, then performing hemolysis treatment and fluorescent reagent treatment on the blood sample, and measuring fluorescent signal information of the blood sample.
In one embodiment of the analysis method, the step 310 may be to measure the blood sample by using flow cytometry to obtain fluorescence signal information, scattered light signal information (forward scattered light signal information or side scattered light signal information), and forward scattered light signal information or side scattered light signal information of the blood sample after the treatment with the fluorescent agent.
The following cell detection can be performed by the acquired fluorescence signal information and scattered light signal information. White blood cell classification and/or enumeration is performed based on the side scatter light values and modified fluorescent signal information of the blood sample, e.g., white blood cell tetra classification via DIFF channels. The nucleated red blood cells are counted based on the forward scattered light value of the blood sample and the corrected fluorescence signal information, for example, through an NRBC channel. White blood cell count and/or reticulocyte identification and/or basophil classification are performed based on the forward scattered light value and the corrected fluorescence signal information of the blood sample, such as reticulocyte classification and/or counting via RET channels. Basophil classification is performed based on the side scatter light values and the corrected fluorescence signal information of the blood sample, for example, BASO channels.
Step 350: and correcting the fluorescent signal information and/or sending out abnormal prompt of the fluorescent signal information according to the red blood cell parameters. And correcting the fluorescence signal information and/or giving out abnormal prompt of the fluorescence signal information according to the red blood cell parameters. The correction can be performed only, or only an abnormality prompt can be sent out, or both correction and abnormality prompt can be performed
In some embodiments, step 350 modifies the fluorescent signal information based on the red blood cell parameter, including: and carrying out increasing correction and/or decreasing correction on the fluorescence signal information according to the red blood cell parameters. The objective of the correction is: regardless of how the HCT, RBC, HGB parameters of the red blood cells change, the fluorescence signal value corresponding to the particle is corrected to be near the normal fluorescence signal intensity value of the particle.
(1) Augmentation correction
Step 350 of performing augmentation correction on the fluorescence signal information according to the red blood cell parameter, including: and if the red blood cell 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 preset value, the greater the red blood cell parameter, the greater the magnitude of the increase correction performed on the fluorescence signal information in step 350. The setting of the first preset value may be based on the normal red blood cell parameter value of the particle under normal fluorescence intensity.
As described above, the red blood cell parameter may comprise 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, a red blood cell parameter greater than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 greater than the first preset value, it means that the red blood cell parameter is greater than the first preset value.
(2) Reduction correction
Step 350 of performing reduction correction on the fluorescence signal information according to the red blood cell parameter includes: 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 preset value, the smaller the red blood cell parameter, the greater the magnitude of the reduction correction performed on the fluorescence signal information in step 350
As described above, the red blood cell parameter may comprise 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, a red blood cell parameter less than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 means that the red blood cell parameter is smaller than the first preset value.
Step 350 performs an increase correction and/or a decrease correction on the fluorescence signal information according to the red blood cell parameter, including: substituting the red blood cell parameter 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 an embodiment, the correction function is a linear function or a power function. In an embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor substitutes the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function, wherein the method comprises the following steps of: substituting the red blood cell parameter as an independent variable into the first function to obtain a correction coefficient, substituting the correction coefficient as a constant and substituting the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information. In an embodiment, the first function and the second function are both linear functions. For example, reference may be made to embodiment 1, and details of the correction function, the first function and the second function will be described in the following embodiments, which are not repeated herein.
Referring to fig. 4, in one embodiment of the analysis method, the method may further include step 370: and outputting the corrected fluorescence signal information, for example, displaying a fluorescence signal scatter diagram through a display according to the corrected fluorescence signal information, or transmitting the corrected fluorescence signal information to a remote server and a central station.
Of course, in some embodiments, after correction, the corrected fluorescence signal information may be output, for example, the corrected fluorescence signal scatter diagram may be displayed through the display according to the corrected fluorescence signal information, and the corrected fluorescence signal scatter diagram may be displayed through the display according to the corrected fluorescence signal information, so as to facilitate comparison by the user.
In some embodiments, after correction, a prompt may also be issued to the user that the fluorescent signal information has been corrected, for example, via a display. In some embodiments, the processor 104 may also output the corrected amplitude of the fluorescence signal information, for example, by displaying the corrected amplitude of the fluorescence signal information in percentage form via a display. In some embodiments, after correction, a prompt or alarm is issued that the fluorescence signal information has been corrected if the magnitude of the correction of the fluorescence signal information exceeds a set threshold.
Example 3
In response to the above-mentioned method for correcting fluorescence signal information of a blood sample, an embodiment of the present invention further discloses a device for correcting fluorescence signal information of a blood sample, referring to fig. 5, the device for correcting fluorescence signal information of a blood sample may include a memory 210 and a processor 230, the memory 210 is used for storing a program, the processor 230 is used for implementing the analysis method of any of the above-mentioned embodiments by executing the program stored in the memory 210, for example, implementing the above-mentioned steps 300, 330 and 350, and may also be used for implementing step 370. Processor 230 may function similarly or identically to processor 104 in a blood cell analyzer. The following is a detailed description.
In one embodiment, processor 230 obtains fluorescence signal information from a blood sample treated with a fluorescent reagent, obtains measured red blood cell parameters from the blood sample, and modifies the fluorescence signal information based on the red blood cell parameters. The fluorescent signal mainly represents white blood cell particles or ghost particles, wherein the ghost particles refer to particles of which the plasma membrane is broken to remain the original form and size of the cell membrane structure after the red blood cells are subjected to hypotonic treatment. In one embodiment, the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.
As described above, the data acquired by the processor 230 includes fluorescence information of the blood sample and red blood cell parameters in the blood sample, and in an embodiment, the processor 230 may also acquire forward scattered light signal information or side scattered light signal information of the blood sample. These three types of data-namely, the first type of data: fluorescence information number information of the blood sample, second kind of data: forward scattered light signal information or side scattered light signal information of the blood sample, data of a third type: one or more of the red blood cell parameters in the blood sample may be transmitted by other devices to processor 230.
Processor 230 corrects the fluorescence signal information based on the red blood cell parameter, which may include: and carrying out increasing correction and/or decreasing correction on the fluorescence signal information according to the red blood cell parameters. The objective of the correction is: regardless of how the HCT, RBC, HGB parameters of the red blood cells change, the fluorescence signal value corresponding to the particle is corrected to be near the normal fluorescence signal intensity value of the particle.
(1) Augmentation correction
Processor 230 performs augmentation correction on the fluorescent signal information based on the red blood cell parameter, including: and if the red blood cell 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 preset value, the greater the red blood cell parameter, the greater the magnitude by which the processor 230 performs the increase correction on the fluorescence signal information. The setting of the first preset value may be based on the normal red blood cell parameter value of the particle under normal fluorescence intensity. As described above, the red blood cell parameter may comprise 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, a red blood cell parameter greater than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 greater than the first preset value, it means that the red blood cell parameter is greater than the first preset value.
(2) Reduction correction
Processor 230 performs a reduction correction on the fluorescent signal information based on 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 less than the first preset value, the smaller the red blood cell parameter, the greater the magnitude of the reduction correction of the fluorescent signal information by the processor 230
As described above, the red blood cell parameter may comprise 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, a red blood cell parameter less than a first preset value may refer to: and (3) carrying out weighted sum or weighted average calculation on two or three of RBC, HGB and HCT included in the red blood cell parameter, wherein a 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 parameter includes three of RBC, HGB, and HCT, the three parameters of RBC, HGB, and HCT are calculated and weighted sum, or weighted average calculation is performed, 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 means that the red blood cell parameter is smaller than the first preset value.
Processor 230 performs an increase correction and/or a decrease correction on the fluorescence signal information based on the red blood cell parameter, including: substituting the red blood cell parameter 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 an embodiment, the correction function may be a linear function or a power function. In an embodiment, the correction function includes a first function and a second function, both of which are increasing functions; the processor substitutes the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function, wherein the method comprises the following steps of: substituting the red blood cell parameter as an independent variable into the first function to obtain a correction coefficient, substituting the correction coefficient as a constant and substituting the fluorescence signal information as an independent variable into the second function to obtain corrected fluorescence signal information. In an embodiment, the first function and the second function are both linear functions. Examples of the correction function, the first function, and the second function may refer to embodiment 1, or refer to specific embodiments described in detail below, and are not described herein.
In one embodiment of the processor 230, the modified fluorescent signal information may also be output, for example, by displaying a fluorescent signal scatter plot on a display according to the modified fluorescent signal information, or by sending the modified fluorescent signal information to a remote server, a central station.
Of course, in some embodiments, after correction, the processor 230 may also output pre-and post-correction fluorescence signal information, e.g., display a post-correction fluorescence signal scatter plot through a display based on the post-correction fluorescence signal information, and display a pre-correction fluorescence signal scatter plot through a display based on the pre-correction fluorescence signal information, for ease of comparison by the user.
In some embodiments, after correction, processor 230 may also issue a prompt to the user that the fluorescent signal information has been corrected, for example, via a display. In some embodiments, the processor 104 may also output the corrected amplitude of the fluorescence signal information, for example, by displaying the corrected amplitude of the fluorescence signal information in percentage form via a display. In some embodiments, after correction, a prompt or alarm is issued that the fluorescence signal information has been corrected if the magnitude of the correction of the fluorescence signal information exceeds a set threshold.
The modification of fluorescence signal information according to the present invention is dependent on red blood cell parameters such as one or more of HCT, RBC and HGB. In the analysis of blood cells, the red blood cell parameters such as HCT, RBC and HGB are conventional measurement parameters, so that the correction of fluorescent signal information can be completed under the conditions of not increasing the consumption of reagents and not increasing the measurement channels, the measurement cost is not increased, and the accuracy of measurement results of measurement channels needing fluorescent 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 illustrates how the present invention can modify the fluorescent signal information based on the red blood cell parameters by way of example.
In the following, the relationship between fluorescence signal information and red blood cell parameters is mainly studied in experiments. Applicant counted 482 samples and plotted a scatter plot with HCT, RBC, HGB of these samples on the horizontal axis and the position of the WBC clusters in the fluorescent direction on the vertical axis, as shown in fig. 6, 7 and 8.
As can be seen from the scatter diagrams of fig. 6-8, the position value of the WBC particle clusters in the fluorescence direction is inversely related to HCT, RBC, HGB, i.e., the smaller HCT, RBC, HGB, the larger the position value of the WBC particle clusters in the fluorescence direction; the larger HCT, RBC, HGB, the smaller the value of the WBC particle clusters' position in the fluorescence direction.
One or more of the HCT, RBC, and HGB may be used to calculate a correction factor, which is then used to correct the fluorescence signal information, e.g., multiplying the correction factor by the fluorescence signal information to obtain corrected fluorescence signal information. Taking fig. 6-8 as an example, the goal of the correction is to make the WBC particle cluster position values in the fluorescence direction substantially constant as a function of HCT, RBC, HGB these parameters. In fig. 6-8, we can target a WBC particle cluster position value equal to 2000 in the fluorescence direction to the target horizon (i.e., the normal fluorescence signal intensity value of the leukocyte particle cluster, which varies from system to system), i.e., the WBC particle cluster position value in the fluorescence direction is maintained substantially near 2000 regardless of changes in these parameters HCT, RBC, HGB. 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 fig. 6, the first preset value corresponding to HCT is in the vicinity of 29; in fig. 7, the first preset value for RBC is around 2.3; in fig. 8, the first preset value corresponding to HGB is around 85. The setting of the first preset value can be understood as a red blood cell parameter value corresponding to the correction coefficient of 1 according to the normal red blood cell parameter value of the particle under the condition of normal fluorescence intensity as a reference.
Taking the fluorescent signal information in the NRBC pass corrected based on HCT as an example, the applicant constructed a first function f based on the fitting result of FIG. 6 11 (x 1 )＝0.038x 1 +0.09, where x 1 The value of HCT is represented, and the correction coefficient can be obtained by substituting the value of HCT into the calculation; as shown in fig. 9, which is a function chart of the first function, the abscissa indicates the value of HCT, and the ordinate indicates the value of the correction coefficient, it can be seen that the larger HCT is, the larger the correction coefficient is, that is, the first function is an increasing function. After calculating the correction coefficient, multiplying the fluorescence information signal by the correction coefficient to obtain corrected fluorescence signal information, e.g. a 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 Representing fluorescence signal information before correction; the correction function is therefore:
y fl ＝f 1 (x 1 ,x fl )＝(0.038x 1 +0.09)*x fl +b；
wherein y is fl Representing the corrected fluorescence signal information; b is a constant, for example 0; the symbols are multipliers.
The first function for correction may correspond to a linear function or may correspond to another functional form, such as a power function. In summary, the correction targets are such that the position value of the WBC particle cluster in the fluorescence direction is maintained substantially near the target line, and the fluorescence signal value corresponding to the particle is corrected to near the normal fluorescence signal intensity value of the particle, regardless of the change of these parameters HCT, RBC, HGB.
A scatter plot of the cell particles (mainly WBC particle clusters in the upper right hand corner and ghost particle clusters in the lower left hand corner) of three samples in the NRBC channel is shown as in fig. 10, where the abscissa is the value of the fluorescence signal information of the cell particles and the ordinate is the forward scattered light signal information of the cell particles. The following is based on the correction function y described above fl ＝f 1 (x 1 ,x fl )＝(0.038x 1 +0.09)*x fl +b makes corrections to the fluorescence signal information for sample 1, sample 2 and sample 3, respectively, where b is taken as 0.
rbc=0.76×10.≡12/L, hgb=26 g/L, hct=8.2% in sample 1. As shown in fig. 10 (a), which shows a scatter diagram of unmodified cell particles obtained in the NRBC channel, it can be seen that the fluorescence signal information of the cell particle mass is large; sample 1 has an HCT value of 8.2% and is taken into the first function f 11 (x 1 )＝0.038x 1 The correction factor was 0.4016 after +0.09. The corrected value of the fluorescence signal information of each cell particle can be obtained by multiplying the correction coefficient 0.4016 by the value of the fluorescence signal information of each cell particle of the sample 1, as shown in fig. 10 (d).
rbc=3.25×10.≡12/L, hgb=71 g/L, hct=23.8% in sample 2. As shown in FIG. 10 (b) which shows a scatter plot of uncorrected cell particles obtained in the NRBC channel, the fluorescence signal of the WBC pellet can be seenThe information is moderate; sample 2 has an HCT value of 23.8% and is taken into the first function f 11 (x 1 )＝0.038x 1 The correction factor was 0.9944 after +0.09. The corrected value of the fluorescence signal information of each cell particle can be obtained by multiplying the correction coefficient 0.9944 by the value of the fluorescence signal information of each cell particle of the sample 2, as shown in fig. 10 (e).
rbc=7.04×10.≡12/L, hgb=158 g/L, hct=50.3% in sample 3. As shown in fig. 10 (c), which shows a scatter diagram of unmodified cell particles obtained in the NRBC channel, the fluorescence signal information of the cell particle mass is small; sample 3 has an HCT value of 50.3% and is taken into the first function f 11 (x 1 )＝0.038x 1 The correction factor was 2.0014 after +0.09. The corrected value of the fluorescence signal information of each cell particle can be obtained by multiplying the correction coefficient 2.0014 by the value of the fluorescence signal information of each cell particle of the sample 3, as shown in fig. 10 (f).
As can be seen from fig. 10 (e) to 10 (f), the fluorescence signals of the cell particle clusters are corrected to a suitable range, that is, to a position relatively offset in the scattergram.
The dye competition of the channel dye method such as NRBC channel, BASO channel, RET channel and DIFF channel causes abnormal fluorescent signals of the leucocyte population, and the classification error of the leucocyte population or NRBC population is easy to cause. In the classification of white blood cells, the applicant has found through long-term research and development that under some samples, when cells are treated with a fluorescent dye, the measurement result deviates from the microscopic examination result when the cells are classified by using a fluorescent signal and a scattered light signal. Through intensive studies, it was found that parameters of erythrocytes in such samples are different from normal values. Through verification, the red blood cell parameter has a certain correlation with the fluorescence signal intensity, and the fluorescence signal can be corrected by using the red blood cell parameter. While not being bound by theory, the applicant speculates that it is possible that erythrocytes will affect the staining behavior of other cells to be classified, resulting in abnormal fluorescent signals.
According to the embodiment of the application, the fluorescent signal is 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 leukocyte classification, taking the fixed demarcation algorithm as an example, the embodiments of the present application can enable the target particle population, such as the leukocyte population, or the nucleated erythrocyte population, or the reticulocyte population, to be accurately located in the preset area of the fixed demarcation algorithm, so as to ensure the accuracy of classification and counting. The application can correct fluorescent signals with red blood cell parameters under NRBC channel, BASO channel, WNB channel, RET channel and DIFF channel so as to correct the position of white blood cell population in a 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 a computer program. When all or part of the functions in the above embodiments are implemented by means of a computer program, the program may be stored in a computer readable storage medium, and the storage medium may include: read-only memory, random access memory, magnetic disk, optical disk, hard disk, etc., and the program is executed by a computer to realize the above-mentioned functions. For example, the program is stored in the memory of the device, and when the program in the memory is executed by the processor, all or part of the functions described above can be realized. In addition, when all or part of the functions in the above embodiments are implemented by means of 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 the program in the above embodiments may be implemented by downloading or copying the program into a memory of a local device or updating a version of a system of the local device, and when the program in the memory is executed by a processor.
The foregoing description of the invention has been presented for purposes of illustration and description, and is not intended to be limiting. Several simple deductions, modifications or substitutions may also be made by a person skilled in the art to which the invention pertains, based on the idea of the invention.

Claims (29)

1. A method of analyzing a blood sample, comprising:
obtaining fluorescent signal information of a blood sample treated by a fluorescent reagent;
obtaining measured parameters of red blood cells in the blood sample; the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration;
and correcting the fluorescent signal information and/or sending out abnormal prompt of the fluorescent signal information according to the red blood cell parameters.
2. The method of analyzing a blood sample according to claim 1, wherein the obtaining fluorescence signal information of the blood sample treated with the fluorescent reagent comprises: and acquiring fluorescence signal information, forward scattering light signal information and/or side scattering light signal information of the blood sample treated by the fluorescent reagent.
3. The method of analyzing a blood sample according to claim 2, wherein the obtaining fluorescence signal information of the blood sample treated with the fluorescent reagent comprises: and measuring the blood sample by adopting flow cytometry to obtain fluorescence signal information, forward scattering light signal information and/or side scattering light signal information of the blood sample treated by the fluorescent reagent.
4. A method of analyzing a blood sample according to claim 2 or 3, further comprising: and performing 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. A method of analyzing a blood sample according to claim 2 or 3, further comprising: and counting nucleated red blood cells according to the forward scattered light value of the blood sample and the corrected fluorescence signal information.
6. A method of analyzing a blood sample according to claim 2 or 3, further comprising: and performing white blood cell count and/or reticulocyte recognition and/or basophil classification according to the forward scattered light value and the corrected fluorescence signal information of the blood sample.
7. A method of analyzing a blood sample according to claim 2 or 3, further comprising: and classifying basophils according to the side scattering light value of the blood sample and the corrected fluorescence signal information.
8. The method for analyzing a blood sample according to claim 1, wherein before obtaining the fluorescence signal information of the blood sample treated with the fluorescent reagent, further comprising: and performing red blood cell parameter measurement on the blood sample to determine the red blood cell parameter, then performing hemolysis treatment and fluorescent reagent treatment on the blood sample, and measuring fluorescent signal information of the blood sample.
9. The method of analyzing a blood sample of claim 8, wherein said performing a red blood cell parameter measurement on said blood sample to determine said red blood cell parameter comprises: and measuring the red blood cell parameters of the blood sample by adopting a small hole impedance method or an optical method to determine the red blood cell parameters.
10. The method of claim 1, wherein said modifying said fluorescent signal information based on said red blood cell parameter comprises: substituting the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as the dependent variables.
11. The method of analyzing a blood sample according to claim 10, wherein the correction function is an increasing function of a dependent variable relative to an independent variable.
12. The method of analyzing a blood sample according to claim 10, wherein the modified function is a linear function, a polynomial function, or a power function.
13. The method of analyzing a blood sample according to claim 1, wherein after correcting the fluorescent signal information according to the red blood cell parameter, further comprising: outputting the corrected fluorescence signal information.
14. The method of claim 13, wherein after correcting the fluorescent signal information based on the red blood cell parameter, further comprising: outputting the fluorescence signal information before and after correction.
15. The method of analyzing a blood sample according to claim 1, wherein after correcting the fluorescent signal information according to the red blood cell parameter, further comprising: and sending out a prompt that the fluorescence signal information is corrected or sending out an alarm.
16. The method of analyzing a blood sample of claim 15, wherein said signaling a notification that the fluorescence signal information has been modified comprises: and outputting the corrected amplitude of the fluorescence signal information.
17. The method of claim 15, wherein after correcting the fluorescent signal information based on the red blood cell parameter, further comprising: and if the corrected amplitude of the fluorescent signal information exceeds the set threshold value, sending a prompt that the fluorescent signal information is corrected or sending an alarm.
18. A blood cell analyzer, comprising:
at least one reaction cell for providing a reaction site for a blood sample and a reagent;
The optical detection device is used for carrying out light irradiation on the blood sample after the reagent treatment, collecting optical signals generated by each particle in the blood sample after the reagent treatment 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 tank to the optical detection device;
the processor is used for receiving and processing the optical signal information output by the optical detection device so as to obtain the measurement parameters of the blood sample; the processor acquires fluorescent signal information of the blood sample treated by the fluorescent reagent, acquires measured red blood cell parameters in the blood sample, corrects the fluorescent signal information according to the red blood cell parameters and/or sends out abnormal prompt of the fluorescent signal information; the red blood cell parameter includes at least one of hematocrit, red blood cell count, and hemoglobin concentration.
19. The blood cell analyzer of claim 18, wherein the processor obtains fluorescence signal information of the blood sample after treatment with the fluorescent reagent, and forward scattered light signal information and/or side scattered light signal information of the leukocyte particles.
20. The blood cell analyzer of claim 19, wherein the optical detection device includes a light source and a flow chamber having an aperture, wherein white blood cell particles in the fluorescent reagent-treated blood sample flow within the flow chamber and pass through the aperture one by one, wherein light emitted from the light source irradiates the particles in the aperture and generates a fluorescent signal and a scattered light signal, respectively, and the fluorescent signal information, and 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.
21. The blood cell analyzer of claim 19 or 20, wherein the processor further performs white blood cell classification and/or counting based on the side scatter light values and the modified fluorescent signal information of the blood sample.
22. The blood cell analyzer of claim 19 or 20, wherein the processor further performs nucleated red blood cell counting based on the forward scattered light values and the corrected fluorescence signal information of the blood sample.
23. A blood cell analyzer according to claim 19 or 20 wherein the processor further performs white blood cell count and/or reticulocyte identification and/or basophil classification based on the forward scattered light values and modified fluorescent signal information of the blood sample.
24. The blood cell analyzer of claim 19 or 20, wherein the processor further performs basophil classification based on the side scatter light values and the modified fluorescent signal information of the blood sample.
25. The blood cell analyzer of claim 18, wherein the reaction cell provides a reaction site for the blood sample and a reagent required for measuring a red blood cell parameter, wherein the optical detection device measures the red blood cell parameter of the blood sample treated by the reagent and outputs optical signal information, and wherein the processor determines the red blood cell parameter based on the optical signal information; and the reaction tank carries out hemolysis treatment and fluorescent reagent treatment on the blood sample, and then the optical detection device measures the treated blood sample and outputs fluorescent signal information of the blood sample.
26. The blood cell analyzer of claim 18, wherein the processor is configured to: substituting the erythrocyte parameter and the fluorescence signal information as independent variables into a preset correction function to obtain corrected fluorescence signal information as the dependent variables.
27. The blood cell analyzer of claim 26, wherein the correction function is an increasing function of the dependent variable relative to the independent variable.
28. The blood cell analyzer of claim 18, wherein the processor further outputs modified fluorescent signal information.
29. A computer readable storage medium storing a program executable by a processor to implement the method of any one of claims 1 to 17.
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