CN114450589A

CN114450589A - Method for analyzing red blood cells in blood sample and blood analysis system

Info

Publication number: CN114450589A
Application number: CN201980100664.6A
Authority: CN
Inventors: 叶波; 祁欢
Original assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Priority date: 2019-10-23
Filing date: 2019-10-23
Publication date: 2022-05-06
Also published as: WO2021077327A1

Abstract

A method of analyzing red blood cells in a blood sample and a blood analysis system (100). The method comprises the following steps: obtaining a first sample from the blood sample, and mixing the first sample with a first reaction reagent to obtain a first suspension, wherein in the first suspension, red blood cells basically maintain the original shape; obtaining a second sample from the blood sample, mixing the second sample with a second reagent comprising a hemolytic agent to obtain a second suspension in which the erythrocytes are lysed; detecting the first suspension to obtain a first electric signal of each cell, and obtaining first integral distribution information of the cells according to the first electric signal; detecting the second suspension to obtain optical information cell by cell, identifying leukocytes through the optical information, and obtaining second volume distribution information of the leukocytes at least according to forward scattered light information of the leukocytes, or detecting the second suspension to obtain a second electrical signal cell by cell and obtaining third volume distribution information of the leukocytes according to the second electrical signal; and correcting the first volume distribution information by using the second volume distribution information or the third volume distribution information to obtain corrected volume distribution information of the red blood cells in the blood sample.

Description

Method for analyzing red blood cells in blood sample and blood analysis system

Technical Field

The invention relates to the field of blood cell detection, in particular to a red blood cell analysis method and an analysis system thereof.

Background

A blood cell analyzer is an instrument that can detect information on cells in blood, and can count and classify cells such as White Blood Cells (WBCs), Red Blood Cells (RBCs), Platelets (PLTs), and the like.

For counting red blood cells and platelets, the most commonly used method is, for example, the pinhole electrical impedance method. In this method, when a cell particle in blood passes through a small hole, a transient voltage change is caused, forming a voltage pulse. The intensity of the voltage pulse reflects the size of the volume of the cells and the number reflects the number of cells passing through the microwells, thereby obtaining a histogram of the volume distribution of the measured blood cells, from which platelets and red blood cells can be distinguished.

The Blood Cell analyzer using the small-hole impedance method can obtain a count value of Red Blood cells, and can obtain commonly used Red Blood Cell parameters such as Mean Red Blood Cell Volume (MCV), Mean Red Blood Cell Hemoglobin content (MCH), Mean Red Blood Cell Hemoglobin Concentration (MCHC), Red Blood Cell Distribution Width Variation Coefficient (RDW-CV), Red Blood Cell Distribution Width Standard Deviation (RDW-SD, Red Blood Cell Distribution Width Standard Deviation), and Hematocrit (HCT, Hematocrit) from histogram Distribution parameters of Red Blood cells.

In practice, however, a large volume of red blood cells may be present in the red blood cells, while some white blood cells, such as small lymphocytes, may also be small in volume, resulting in the two not being well differentiated on the histogram.

Under normal conditions, the number of white blood cells is much lower than red blood cells, and the difference between the numbers is about 1000 times. Therefore, the influence of a small amount of white blood cells on the counting of red blood cells, the measurement of parameters such as hemoglobin content HGB and MCV is almost negligible. However, it has been reported that when the amount of leukocytes exceeds 40.0X 10^9/L, the leukocytes affect the analysis of hemoglobin content and also falsely increase morphological parameters such as red blood cell count and MCV. Finally, the result of calculating the red blood cell count value and the related parameters is inaccurate.

In order to obtain more accurate red blood cell parameters, a whole blood sample can be added into a hematocrit tube for low-speed centrifugation, a leucocyte layer and a plasma layer are removed, and then an equal amount of diluent is added and mixed uniformly, and then an analyzer is used for detection. This obviously increases the complexity of the test and does not satisfy the requirement of rapid analysis of blood.

Furthermore, Zezang et al (test medicine, 2010,25(3):244-246) also reported that abnormal increase in leukemia caused by certain diseases (such as leukemia) interfered with the red blood cell parameters of all models of hematology analyzers, and provided a simple correction method. The method directly subtracts the white blood cell count from the raw red blood cell count to obtain a corrected red blood cell count. However, this method can only obtain the count of the red blood cells, but cannot obtain all morphological parameters of the red blood cells, and the practical application range is too narrow.

Therefore, a method and an analyzer capable of rapidly and accurately acquiring RBC count values and morphological parameters are needed.

Disclosure of Invention

Aiming at the defect that the counting result and the morphological parameter measuring result of the red blood cells of the existing blood cell analyzer which measures the red blood cells by the electrical impedance principle are easy to interfere when detecting a high-value white blood cell sample, the invention provides an analyzing method for removing the interference of the white blood cells on the red blood cells and a corresponding blood cell analyzer.

According to a first aspect of the present invention, there is provided a method of analyzing red blood cells in a sample. The method comprises the following steps:

obtaining a first sample from the blood sample, and mixing the first sample with a first reaction reagent to obtain a first suspension, wherein in the first suspension, red blood cells basically maintain the original shape;

obtaining a second sample from said blood sample, and mixing said second sample with a second reagent comprising a hemolysing agent to obtain a second suspension, wherein red blood cells are lysed in said second suspension;

detecting the first suspension to obtain a first electric signal of each cell, and obtaining first integral distribution information of the cells according to the first electric signal;

detecting said second suspension to obtain cell-by-cell optical information, identifying leukocytes from said optical information, obtaining second volumetric distribution information of leukocytes from at least forward scattered light information of said leukocytes, or

Detecting the second suspension to obtain a second electric signal of each cell, and obtaining third volume distribution information of white blood cells according to the second electric signal;

and correcting the first volume distribution information by using the second volume distribution information or the third volume distribution information to obtain corrected volume distribution information of the red blood cells in the blood sample.

According to a second aspect of the present invention, a blood analysis system is provided. The blood analysis system includes: sampling portion, reagent supply unit, reaction unit, detection portion, controller and processor, wherein:

the sampling part is configured to obtain at least two samples from a blood sample and respectively convey the samples to the reaction part;

the reagent supply part configured to supply a required reaction reagent to the reaction part;

the reaction part comprises a first reaction chamber and is used for mixing a first sample and a first reaction reagent to prepare a first suspension; and a second reaction chamber for mixing a second sample with a second reaction reagent to prepare a second suspension, and the reaction part is configured to supply the first suspension and the second suspension to the detection part, respectively;

the detection part comprises an electrical detection part and an optical detection part, wherein the electrical detection part is configured to enable the particles in the first suspension to pass through the resistance detector one by one to obtain a first electric signal of the particles in the first suspension, and the optical detection part is configured to enable the particles in the second suspension to pass through the optical detector one by one to obtain optical information of the particles in the second suspension;

the controller is coupled with the sampling part, the reagent supply part, the reaction part and the detection part and controls the actions of the sampling part, the reagent supply part, the reaction part and the detection part; and

the processor, the processor and the detection part are coupled, wherein

The processor acquires the electrical signal and the optical information from the detection portion, acquires first volume distribution information from the electrical signal, acquires white blood cell distribution information from the optical information, acquires second volume distribution information using forward scattered light intensity, and corrects the first volume distribution information with the second volume distribution information to acquire corrected volume distribution information of red blood cells in the blood sample.

According to a third aspect of the present invention, a further blood analysis system is provided. The blood analysis system includes:

sampling portion, reagent supply portion, reaction unit, detection portion and controller, wherein:

a sampling section configured to take at least two samples from a blood sample and transfer the samples to the reaction section, respectively;

a reagent supply portion configured to supply a required reaction reagent to the reaction portion;

a reaction part including a first reaction chamber for mixing a first sample with a first reaction reagent to prepare a first suspension, and a second reaction chamber for mixing a second sample with a second reaction reagent to prepare a second suspension, and configured to supply the first suspension and the second suspension to the detection part, respectively;

a detection portion comprising an electrical detection portion, wherein the electrical detection portion is configured to pass particles in the suspension one by one through a resistance detector to obtain a first electrical signal of a first suspension and a second electrical signal of a second suspension;

a controller coupled to the sampling part, the reagent supplying part, the reaction part and the detection part, for controlling the operations of the sampling part, the reagent supplying part, the reaction part and the detection part; and

a processor coupled with the detection portion, wherein

The processor obtains the first and second electrical signals from the detection portion, obtains first volume distribution information from the first electrical signal, obtains third volume distribution information from the second electrical signal, and corrects the first volume distribution information with the third volume distribution information to obtain corrected volume distribution information of red blood cells of the blood sample.

According to a fourth aspect of the present invention, there is provided a computer readable storage medium having a computer program stored thereon, the computer program when executed by a processor implementing the steps of:

acquiring a first electric signal obtained by detecting a first suspension obtained by mixing a first sample acquired from a blood sample with a first reaction reagent, and obtaining first integrated distribution information from the first electric signal, the electric signal being obtained by detecting the first suspension by an electrical detection section;

acquiring optical information obtained by detecting a second suspension obtained by mixing a second sample obtained from the blood sample with a second reaction reagent containing at least a hemolytic agent, and obtaining leukocyte distribution information from the optical information, and obtaining second volumetric distribution information using a forward scattered light intensity,

or

Acquiring a second electric signal obtained by detecting a second suspension obtained by mixing a second sample obtained from a blood sample with a second reaction reagent containing at least a hemolytic agent, and obtaining third volume distribution information from the second electric signal, the electric signal being obtained by detecting the second suspension by the electric detection portion; and

The blood cell analysis method of the invention can effectively eliminate the interference of the white blood cells on the red blood cell analysis in the sample, especially the sample with higher content of white blood cells, by utilizing the existing white blood cell (hemolysis) optical detection channel or only utilizing the white blood cell (hemolysis) electrical impedance detection channel, so as to obtain accurate red blood cell count and related parameters, thereby providing accurate detection information for clinical diagnosis and treatment. In a conventional blood analyzer, white blood cells are detected in a WBC/BASO channel, a DIFF channel, an NRBC channel, and an electrical impedance detection channel, and a count value is obtained. It is confirmed by the examples detailed below that accurate red blood cell volume distribution information can be obtained regardless of which channel the white blood cell volume distribution information obtained by the electrical impedance method is used to correct the volume distribution information including white blood cells and red blood cells obtained by the electrical impedance method. In addition, in the leukocyte (hemolysis) electrical impedance detection channel, leukocyte volume distribution information can also be obtained through the detection of the hemolysis sample, and the leukocyte volume distribution information is used for correcting the volume distribution information containing leukocytes and erythrocytes to obtain accurate erythrocyte volume distribution information. The method and the analysis system of the invention can obtain accurate red blood cell count and related morphological parameters in a simple method.

Drawings

FIG. 1A is a schematic flow chart of the method steps for analyzing red blood cells in a blood sample according to a first embodiment of the present invention;

FIG. 1B is a schematic flow chart of the method steps for analyzing red blood cells in a blood sample according to a second embodiment of the present invention;

FIG. 2A is a block diagram schematically illustrating the structure of a blood analysis system according to a first embodiment of the present invention;

FIG. 2B is a block diagram schematically showing the structure of a blood analysis system according to a second embodiment of the present invention;

FIG. 3 is a first schematic diagram of a first test solution according to example 1;

FIG. 4 is a scattergram of leukocyte populations of the second test solution according to example 1;

FIG. 5 is an overlay of a first histogram and a second histogram according to example 1, wherein A is a full graph and B is an enlarged view of the 100-300fL region;

FIG. 6 is a corrected red blood cell histogram according to example 1;

FIG. 7 is a first schematic diagram of a first test solution according to example 2;

FIG. 8 is a scattergram of leukocyte populations of the second test solution according to example 2;

FIG. 9 is an overlay of a first histogram and a second histogram according to example 2, wherein A is a full graph and B is an enlarged view of the 100-300fL region;

FIG. 10 is a corrected red blood cell histogram according to example 2;

FIG. 11 is a first schematic diagram of a first test solution according to example 3;

FIG. 12 is a scattergram of leukocyte populations of the second test solution according to example 3;

FIG. 13 is an overlay of a first histogram and a second histogram according to embodiment 3, wherein A is a full graph and B is an enlarged view of the 100-300fL region;

FIG. 14 is a corrected red blood cell histogram according to example 3;

FIG. 15 is a first schematic diagram of a first test solution according to example 4;

FIG. 16 is a scattergram of leukocyte populations of the second test solution according to example 4;

FIG. 17 is an overlay of a first histogram and a second histogram according to example 4, wherein A is a full graph and B is an enlarged graph of the area of 150 and 300 fL;

FIG. 18 is a corrected red blood cell histogram according to example 4;

FIG. 19 is a first schematic diagram of a first test solution according to example 5;

FIG. 20 is a second histogram of leukocyte populations of the second test solution according to example 5;

FIG. 21 is an overlay of a first histogram and a second histogram according to example 5, wherein A is a full graph and B is an enlarged view of the area of 100 and 200 fL;

FIG. 22 is a corrected red blood cell histogram according to example 5; and

fig. 23 is a histogram of red blood cells obtained according to the direct test of comparative example 1.

Detailed Description

The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the invention are shown. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present definition takes precedence.

The term "histogram" as referred to in this context is a graphical form of the cell volume distribution, a common form presenting a continuously variable probability distribution. Alternatively, the cell volume distribution may be presented in numerical form as a table or list having equal or similar resolution to the volume histogram, or in any other suitable manner known in the art.

Aiming at the problem that the analysis of the current blood analyzer aiming at a high white blood cell sample has interference on red blood cell counting and morphological parameters, the invention provides a simple and convenient red blood cell analysis method for removing the interference of white blood cells.

For normal samples, the white blood cell content is only one thousandth of the red blood cells, and the volume of white blood cells is typically larger than red blood cells. In the histogram obtained by the conventional impedance method, the influence of white blood cells is negligible. However, in the case of abnormal samples with high leukocyte content, the histogram obtained by the conventional impedance method contains a considerable amount of leukocyte particles mixed in the volume range of the red blood cells, thereby affecting the counting of the red blood cells and other morphological analysis results, and further adversely affecting the correct diagnosis.

In general, the invention analyzes and obtains the histogram of the white blood cells by utilizing the existing channel and the detection equipment in the blood analyzer, and corrects the red blood cell histogram detected according to the conventional impedance method according to the histogram of the white blood cells, thereby obtaining the corrected histogram which is close to the real red blood cell information, and further obtaining the correct counting of the red blood cells and other morphological analysis results.

A method of analyzing red blood cells in a blood sample according to a first embodiment of the present invention comprises the steps of:

obtaining a first sample from said blood sample, mixing said first sample with a first reactive reagent to obtain a first suspension, and obtaining a second sample from said blood sample, mixing said second sample with a second reactive reagent comprising a hemolysing agent to obtain a second suspension, wherein in said first suspension red blood cells substantially maintain their original morphology and in said second suspension red blood cells are lysed;

detecting the first suspension to obtain a first electric signal of each cell, and obtaining first integral distribution information according to the first electric signal;

detecting the second suspension to obtain optical information of each cell, obtaining leukocyte distribution information through the optical information, and obtaining second volume distribution information at least according to the forward scattering light information of the leukocytes;

and correcting the first volume distribution information by using the second volume distribution information to obtain corrected volume distribution information of the red blood cells in the blood sample.

Referring to fig. 1A, a schematic flow diagram of the method of this embodiment is illustrated. In this embodiment, a blood sample is first provided S11. The blood sample of the present invention is typically a whole blood sample. Can be a whole blood sample from a mammal, particularly a primate, more particularly a human.

Obtaining a first specimen from said blood sample is mixed with a first reaction reagent S22 to obtain a first suspension.

The first reactive agent is typically a diluent. The diluent is not particularly limited in the present invention, and any diluent suitable for electrical impedance detection may be used in the present invention. The diluent typically includes one or more salts, such as an alkali metal salt, and is adjusted to be isotonic (isotonics) to maintain red blood cell volume. The diluent is not particularly limited in the present invention, and any diluent suitable for electrical impedance detection may be used in the present invention. For example, any suitable commercial blood diluent can be used, such as M-68DS diluent, M-53D diluent, etc., from Shenzhen Meyer biomedical electronics, Inc. (Shenzhen, China).

The first suspension obtained by dilution, i.e., the first test solution, is further subjected to electrical impedance detection S23 to obtain an electrical signal of blood cells in the first sample. The electrical impedance method may be, for example, a pinhole impedance method, a sheath flow impedance method, or the like.

First, electrical information of the whole particle events in the first sample is acquired, and further, volume distribution information of blood cells (referred to as a first histogram) is obtained therefrom S24.

In other embodiments, red blood cells and platelets can be distinguished, with the red blood cell histogram as the first histogram. Simultaneously, or sequentially, a second sample is taken from the blood sample and mixed with a second reactive reagent S32 to obtain a second suspension.

In an alternative, the first sample may be sequentially subjected to electrical impedance detection and optical detection. In this case, the second sample is obtained by mixing the first sample subjected to the electrical impedance detection with a second detection reagent. That is, the first blood sample is mixed with a first detection reagent to obtain a first sample, and the first sample is further mixed with a second detection reagent after electrical impedance detection to obtain a second sample.

The benefit of this alternative is that no additional sample size is required, and therefore no additional burden is placed on the subject, particularly those with blood draw difficulties.

The second reaction reagent of the present invention contains a hemolytic agent for lysing erythrocytes in the second sample. The hemolytic agent can be any one or combination of cationic, nonionic, anionic and amphiphilic surfactants. The hemolytic agent for lysing erythrocytes in the second sample is not particularly limited in the present invention. Thus, any lysis reagent suitable for use in the white blood cell classification of a hematology analyzer may be used. The lytic reagent used for the leucocyte differentiation of a hematology analyzer is generally an aqueous solution containing one or more of the above-mentioned hemolytic agents.

In some embodiments, the second reagent for processing the second sample further comprises a fluorescent dye for staining the nucleated blood cells in addition to the hemolytic agent, so that the leukocytes can be further separated and/or distinguished by detecting the intensities of the scattered light and the fluorescence. For example, dissolving reagent formulations described in U.S. patent No. 8,367,358, the entire disclosure of which is incorporated herein by reference, may be employed. The lytic reagent disclosed in U.S. patent No. 8,367,358, which comprises a cationic cyanine compound (a fluorescent dye), a cationic surfactant, a nonionic surfactant and an anionic compound, can be used to lyse red blood cells and classify white blood cells into their subpopulations using fluorescence and light scattering measurements. Other suitable fluorescent dyes may also be used in the lysis reagent. For example, fluorescent dyes are described in U.S. patent No. 8,273,329, the entire disclosure of which is incorporated herein by reference.

In addition, in some embodiments, the fluorescent dye may be included in a separate staining solution as a third reactive agent for use with a second reactive agent that is a lysing agent that does not contain the fluorescent dye. The staining solution may be added to the blood sample before, after, or simultaneously with the lysing agent to stain the nucleated blood cells.

Still referring to fig. 1A, in step S33, the second suspension is further optically detected to obtain optical information of the blood cells in the second suspension.

In step S34, optical information of the cells in the second suspension can be obtained from the optical information; the leukocyte particle population is then differentiated based on the optical properties of the particles, and a histogram of the leukocyte volume is obtained as a second histogram based further on the intensity of the forward scattered light of the leukocytes, which reflects cell volume information.

Obtaining a scatter plot from the optical information in order to identify the population of white blood cells may employ any conventional method. The two-dimensional scattergram is generally obtained by two kinds of optical information such as light intensity of forward and side scattered light, light intensity of forward scattered light and fluorescent light, light intensity of side scattered light and fluorescent light, and the like. If necessary, the three-dimensional scattergram can be obtained through the three kinds of information. The present invention is not particularly limited in this regard.

After the white blood cell particle group is identified, the volume distribution information of the white blood cells can be obtained by analysis according to the intensity of the forward scattered light of the white blood cells and can be represented in a form of a histogram, for example.

The method for converting the optical information into the histogram is not particularly limited, and may be any suitable method that is available, for example, a one-dimensional volume distribution of particles, i.e., a histogram, is obtained by calculation using a forward light scattering signal according to Mie scattering theory.

In addition, any of the following methods may be used for the histogram of the scattergram in the present invention.

One method is to calculate the derivative volume Vol of each particle according to the following equation (1) using the forward light scattering information of the particles_f1。

Vol _f1＝α×FSC (1)

The FSC is the intensity of forward scattered light of a single event (individual event) in a particle swarm to be converted (specifically, a single leukocyte) in a two-dimensional scattergram, α is a constant, the value of α is related to detection equipment, and α values of different detection equipment can be different.

Another method is to calculate the derivative volume Vol of each particle according to equation (2) using the forward light scattering information of the particle_f2。

Vol _f2＝β×exp(γ·FSC) (2)

Where FSC is also the intensity of forward scattered light of a single event (individual white blood cell in the present invention) in the population of particles to be transformed in the two-dimensional scattergram, and β and γ are constants. Similarly, the values are related to the detection devices, and the beta and gamma values can be different for different detection devices.

By converting the light intensity of the forward scattered light of each particle of the second population of particles into a derivative volume Vol_f1Or Vol_f2A volume distribution histogram relating to the white blood cell volume in the histogram obtained by the DC impedance method can be obtained.

The inventors have found that a histogram obtained by electrical impedance detection can be well matched by performing a histogram of a leukocyte scatter diagram obtained by optical detection. Therefore, the invention can use the white blood cell information obtained by optical detection to correct the mixed information of the red blood cells and the white blood cells obtained by electrical impedance detection, and further obtain accurate red blood cell information. Moreover, it has been verified that in this embodiment, whatever channel (such as WBC/BASO, DIFF or NRBC) is used for the leukocyte optical information obtained by leukocyte detection, after conversion into a histogram, can be used to correct the histogram obtained by electrical impedance spectroscopy. The following examples also demonstrate that the corrected red blood cell histograms obtained are consistent with those obtained from a blood sample previously treated to remove leukocytes and then examined by electrical impedance.

Unlike some simple prior art correction methods by subtracting the white blood cell count, the method of the present invention can obtain the volume distribution information of the red blood cells, and thus further obtain the relevant red blood cell morphological distribution data, without being limited to the correction of the red blood cell count.

As shown in fig. 1A, the histogram obtained after correcting the first histogram with the second histogram (step S15) may obtain the red blood cell count and other morphological parameters (step S16). The other morphological parameter may be at least one parameter selected from the group of: mean Corpuscular Volume (MCV), mean corpuscular hemoglobin content (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), coefficient of variation of distribution width of erythrocytes (RDW-CV), standard deviation of distribution width of erythrocytes (RDW-SD), Hematocrit (HCT).

In addition, the method of the invention does not change the detection principle of the signal and can be applied to various detection occasions. That is, the method of the present invention can be used in a scenario where a sample is detected after various hemolyzations in a blood analyzer. Thus, in the case of blood sample testing with a hematology analyzer, any desired other parameters of the blood sample conventionally obtainable by electrical impedance and/or optical testing may be obtained while obtaining the correct red blood cell related parameters. For example: basophils and other leukocyte granules can be distinguished according to the intensity of light scattered by lateral and forward light; basophils and other leukocyte granules can be distinguished according to the light intensity and fluorescence intensity of forward light scattering, and blood cells with nucleic acid substances, such as nucleated red blood cells, can be identified; and four classification of leukocytes according to the intensity of light scattered from the side light and the intensity of fluorescence, distinguishing eosinophils, neutrophils, monocytes, and lymphocytes, but not limited thereto.

After obtaining the relevant information and parameters, the method of the present invention further comprises the step of outputting the required information and/or parameters. The output corrected information (such as histogram) and/or parameters (such as red blood cell count and other morphological parameters) may be further indicated according to whether the white blood cell count is abnormal (e.g. exceeds a first threshold), or according to whether the difference between the red blood cell counts before and after correction exceeds a normal range (e.g. exceeds a second threshold).

The method of the present invention can output not only the corrected histogram and the relevant parameters but also the first histogram before correction, or even the first histogram on which the second or third histogram for correction is superimposed, as necessary.

In addition to outputting the detection result, the normal range of each item may be further output as a reference.

With further reference to FIG. 2A, there is shown a schematic diagram of a blood analyzer 100 that can be used in the above-described method.

The blood analyzer 100 conventionally includes a sampling part 110, a reagent supplying part 120, a reaction part 130, a detection part 140, a controller 150, and a processor 160. Of course, other necessary devices, such as a display, an input and output, a housing, and necessary pumps and fluid paths, etc., may be included, and will not be described in detail herein.

The sampling part 110 may include a pipette device, a sample splitter, etc. to suck the first and second samples for detection from the blood sample and supply the first sample to the first reaction chamber 131 of the reaction part 130 and the second sample to the second reaction chamber 132.

The reagent supply part 120 is used to supply a desired reaction reagent to the reaction part. FIG. 2A shows a reagent supply 120 having two reservoirs according to one embodiment: the reservoirs a121 and B122 are used for storing the first reactive reagent and the second reactive reagent, respectively, as described above, and supplying them to the first reaction chamber 131 of the reaction portion 130, respectively, and supplying the second sample to the second reaction chamber 132 to be mixed with the first and second samples therein, respectively, to obtain a first suspension and a second suspension. The reagent supply 120 may also have more reservoirs. For example, in the embodiment of fluorescent staining of cells as described above, the reagent reservoir section 120 may further include a third reservoir for separately storing the fluorescent staining solution.

In other embodiments, the reagent supply may not have a reservoir. The reagent supplying part sucks and supplies a required reaction reagent from a reagent tank outside the machine to the reaction part.

According to one embodiment, the reaction portion 130 may have a plurality of reaction chambers (as shown, two reaction chambers: a first reaction chamber 131 and a second reaction chamber 132 are shown), and different samples to be tested may be respectively provided to the different reaction chambers to be mixed with corresponding reaction reagents. According to another embodiment, the first reaction chamber and the second reaction chamber in the reaction portion 130 may be a single reaction chamber, and the sample to be tested is sequentially supplied to the reaction chamber to be mixed with the corresponding reaction reagent and then supplied to the detection portion 140 for detection, or the mixture is further added to and mixed in the single reaction chamber. The specific structure of the reaction part 130 may be determined according to actual needs.

After the completion of the mixing reaction, the first suspension and the second suspension are supplied to the electrical impedance detection section 141 and the optical detection section 142 of the detection section 140, respectively, in this embodiment.

The electrical impedance detection section 141 is conventionally equipped with a DC impedance detector and a flow path of a non-focused flow orifice or a focused flow orifice. When particles or blood cells suspended in a conductive solution pass through an orifice provided on a flow path, an electrical signal can be measured based on a change in impedance. The pulse shape, height and width of the impedance signal are directly related to the size or volume of the particle and can be converted to the volume of the particle. When two or more particles having different sizes are measured, the size distribution of these particles may be reflected by the frequency histogram of the impedance signal (e.g., embodied in the form of a histogram). Methods of detecting blood cells by a blood analyzer equipped with a DC impedance measurement device are known, such as described in U.S. Pat. nos. 2,656,508 and 3,810,011, the entire disclosures of which are incorporated herein by reference.

The optical detection section 142 may conventionally include a light source, an optical flow cell, and at least one optical detector. The optical flow cell is a flow-through cell (focused-flow cell) adapted to detect a focused flow of light scattering signals and fluorescence signals. Optical flow cells such as those used in existing flow cytometers and hematology analyzers can be used in the blood analysis system of the present invention. When a particle passes through a detection aperture (orifice) of an optical flow cell, an incident light beam from a light source impinges on the particle passing through the detection aperture, scattering in all directions. For particles stained with a fluorescent stain, fluorescence may also be emitted. Thus, optical detectors may be arranged at different angles relative to the incident light beam to capture the scattered and/or fluorescent light. Since different particle groups have different light scattering characteristics or different fluorescence characteristics according to different fluorescent dyes, it is possible to distinguish different particle groups according to these optical information. In some embodiments, the detector may be arranged at an angle of about 1 ° to about 10 ° relative to the incident light beam to capture the signal of forward light scattering. In other embodiments, the signal of forward light scattering may be collected at an angle of about 2 ° to about 6 ° to the incident light beam. The optical detector is arranged at an angle of about 90 ° with respect to the incident light beam to capture the signal of the side light scatter. The fluorescence detector may also be disposed at an angle of about 90 deg. with respect to the incident beam. In some embodiments, the optical detector that captures the signal of side light scattering can also be arranged at a high angle of about 65 ° to about 115 ° with respect to the incident light beam.

The electrical impedance detection part 141 and the optical detection part 142 that can be used in this embodiment of the present invention may be any suitable known devices, and thus, will not be described herein again.

The blood analysis system of the present invention also has a controller 150. The controller 150 is coupled to the sampling unit, the reagent supply unit, the reaction unit, and the detection system, and controls operations of the sampling unit, the reagent supply unit, the reaction unit, and the detection system.

The blood analysis system of the present invention also has a processor 160. The processor may execute a computer program stored in a memory (not shown) to perform the analysis of the blood sample analysis method described above to obtain corrected red blood cell volume distribution information (histogram) and further to obtain red blood cell count and other morphological results, as well as analysis of other particles in the blood and to obtain related results.

Furthermore, the blood analysis system of the present invention may also have a user interface (not shown). Conventionally, a user interface may include an input device and an output device. Wherein the output device is used for outputting the detection result obtained by the method according to the invention and comprising the graph and the data form. According to other embodiments, the output device can also output the normal value reference ranges of the parameters at the same time, and mark abnormal conditions. In addition, as described above, the output detection result may be corrected to have a certain difference from that before the correction, and may be indicated.

The blood analysis system of the present invention may also have a storage device (not shown). According to other embodiments, the storage device may also be connected to the blood analysis system of the present invention by an external device. The storage devices may store the basic programs and data structures for implementing the functions of the various aspects of the methods described herein. Conventionally, a storage device may include one or more memories and one or more non-transitory computer-readable storage media. The non-transitory computer readable storage medium may include a hard drive, a floppy disk, a compact disk, a secure digital memory card (SD card), a flash memory, and the like. The memory may include main Random Access Memory (RAM) or dynamic RAM (dram) for storing program treatments and data, and Read Only Memory (ROM) for storing fixed instructions. The non-transitory computer readable medium is programmed by a computer application to implement the functions disclosed herein and the corresponding program is executed by one or more processors. When the processor executes a computer application stored on a non-transitory computer readable medium, the processor corrects the first volumetric distribution information and further obtains a red blood cell count and associated morphological parameters according to the methods disclosed herein.

A second embodiment of the present invention is further explained with reference to fig. 1B and 2B. Wherein a schematic flow chart of the steps of the second embodiment is shown in FIG. 1B; fig. 2B shows a schematic configuration diagram of the blood analysis system used in this embodiment.

This second embodiment differs from the first embodiment described above mainly in that the second suspension obtained after the second sample is mixed with the second reagent containing the hemolytic agent (step S42) is also subjected to electrical impedance detection (step S43), so that a third histogram on white blood cells can be obtained based on the second threshold value (step S44).

The corresponding blood analysis system 200 for this embodiment may have substantially the same sampling part 210, reagent supply part 220 (shown with reservoir a221 and reservoir B222), reaction part 230 (shown with first reaction chamber 231 and second reaction chamber 232), and controller 250 as blood analysis system 100, except that detection part 240 may include only electrical impedance detection part 241.

According to this embodiment, after hemolyzing the red blood cells in the second sample, detection is also performed by the electrical impedance method. Since the hemolytic agent may shrink the other blood cells to some extent in addition to lysing the red blood cells, the white blood cell volume value obtained by the second electric signal obtained in step S43 and the first electric signal obtained in step S23 may not be consistent. The inventor of the present invention has found that, the trend of the cell volume change is related to the hemolytic agent used, the volume adjustment coefficient K can be obtained through statistics, and a functional mapping relationship exists between the leukocyte volume obtained by the first electrical signal analysis and the leukocyte volume obtained by the second electrical signal analysis:

VOL2＝K×VOL1 (3)。

the coefficient K can be obtained by performing the same processing and detection as in this embodiment using a sample containing only leukocytes as the first test sample and the second test sample, respectively. The volumes corresponding to the peaks of the two histograms are compared, or the mean cell volumes of the two are calculated separately and compared to obtain the value of K.

The volume distribution obtained by the second electrical signal is corrected according to the K value, and the volume distribution information of the full particle event (i.e. third volume distribution information, such as a third histogram) is obtained.

Furthermore, the coefficient K' can also be obtained directly from the ratio of the leukocyte electrical signals (e.g. the average electrical signal) obtained from the non-hemolytic channel and the hemolytic channel:

S2＝K’×S1 (4)。

from K', the second electrical signal can be directly corrected and a third histogram can be obtained from the corrected second electrical signal.

Next, in step S17, a corrected red blood cell volume distribution histogram is obtained by correcting the first histogram with the third histogram, and further, in step S18, the count of red blood cells and other morphological distribution results are obtained.

When this embodiment is employed,

processors

160, 260 may perform the analysis steps described above to obtain the first histogram and the third histogram, respectively, and perform the correction to obtain a corrected volume distribution histogram of red blood cells, thereby further obtaining red blood cell counts and other morphological distribution results.

The steps, reagents, components and the like relating to this embodiment which are the same as those in the first embodiment will not be described in detail herein.

Furthermore, the invention further provides a computer-readable storage medium. The computer-readable storage medium has stored thereon a computer program which, when being executed by the processor, carries out the steps of the aforementioned blood sample analysis method. The computer readable storage medium may be Memory such as FRAM, ROM, PROM, EPROM, EEPROM, Flash Memory, magnetic surface Memory, optical disk, or CD-ROM.

The invention and its advantageous effects are further illustrated by the following specific examples.

The following examples all measured and analyzed whole blood samples using a commercial blood analyzer BC-6800 (Shenzhen Merrier biomedical electronics, Inc., Shenzhen, China).

The BC-6800 hematology analyzer includes a complete blood cell technology (CBC) module and a classification module. The CBC module includes a first mixing chamber and a DC impedance detector. The mixing chamber is configured to mix a sample taken from a blood sample with a diluent to form a first test solution, and the DC impedance detector is configured to measure a DC impedance signal of the first test solution flowing through an aperture provided in the flow path. The sorting module includes a second mixing chamber, an infrared semiconductor laser, and a plurality of optical detectors. The mixing chamber is configured to mix another aliquot taken from the same blood sample with a hemolysing agent and optionally a fluorescent dye to form a second test solution. The infrared semiconductor laser is used as a light source and emits a light beam which is aligned with the detection hole of the optical flow chamber. The plurality of optical detectors includes a forward scatter light detector capable of detecting a forward scatter signal emitted from the detection well at an angle of about 1 ° to about 10 ° from an incident light beam of the light source, a side scatter light detector capable of detecting a side scatter signal emitted from the detection well at an angle of about 65 ° to about 115 ° from the incident light beam, and a fluorescence detector capable of detecting a fluorescence signal.

Other existing hematology analyzers are also suitable for use with the presently disclosed solution, provided that these detectors are capable of detecting forward scattered light, side scattered light, and/or fluorescent signals. Instead of the side scattered light, and in combination with other light signals, the white blood cells and other cells may be distinguished, and then a white blood cell volume distribution histogram may be obtained based on the forward scattered light of the white blood cells.

Example 1: analysis of red blood cells in a sample using BASO channels

In the CBC module, 4 μ L of anticoagulated whole blood sample was mixed with 1.5mL of M-68DS diluent (Shenzhen Meyer biomedical electronics, Inc.) to form a first sample solution. The first sample solution is sent to a DC impedance detector, an impedance electrical signal is detected, and a first histogram (in which the particles included are denoted as a particle group 1, see fig. 3) is obtained by analysis.

In the sorting module, 20 μ L of an anticoagulated whole blood sample taken from the same blood sample was mixed with 1mL of M-68LD Lyse to Lyse the red blood cells, forming a second reagent solution. The M-68LD Lyse is an aqueous solution containing a cationic surfactant, a nonionic surfactant and an anionic compound, and is used for lysing erythrocytes in a blood sample. The second sample solution is sent to an optical detection system, and a scattered light detector detects the forward scattered light and the side scattered light, and analyzes the light intensities of the obtained forward and side scattered light, thereby distinguishing the white blood cell population from other cells (denoted as a population 2), as shown in a scattergram in fig. 4. The forward scattered light intensity of each cell in the leukocyte population is calculated using the aforementioned formula (1) to obtain a derived volume for each cell, thereby obtaining a second histogram.

The first histogram is overlapped with the second histogram, as shown in FIG. 5A, wherein the curve of the particle group 2 under the curve of the particle group 1 can be clearly seen from the enlarged view of the area of 100 and 300fL (see FIG. 5B). The two curves do not coincide, indicating that red blood cells and white blood cells are mixed in this region of the particle group 1.

By calculation, the histogram of the particle group 2 is subtracted from the histogram of the particle group 1, obtaining a histogram of a corrected population of red blood cells (see fig. 6).

Example 2: analysis of red blood cells in a sample using BASO channels

Another blood sample was tested in the same manner as in example 1. First, a first test solution is detected in the CBC module to obtain a first histogram (particles included therein are denoted as a particle group 1, see fig. 7).

Next, the second sample solution is detected in the classification module to obtain forward scattered light and side scattered light, and the light intensities of the obtained forward and side scattered light are analyzed to distinguish the white blood cell population from other cells (denoted as a population 2), as shown in the scattergram of fig. 8. The forward scattered light intensity of each cell in the leukocyte population is calculated using the aforementioned formula (1) to obtain a derived volume for each cell, thereby obtaining a second histogram.

The first histogram is overlapped with the second histogram, as shown in fig. 9A, in which an enlarged view of the area of 100 and 300fL (see fig. 9B) clearly shows the curve of the particle group 2 below the curve of the particle group 1.

As can be seen in fig. 9, the white blood cells showed two peaks in the impedance channel. As can be seen, the two leukocyte populations interfere with the population of red blood cells. The corrected red blood cell histogram is shown in fig. 10.

Example 3: analysis of red blood cells in a sample using a DIFF channel

In the CBC module, 4 μ L of anticoagulated whole blood sample taken from one blood sample was mixed with 1.5mL of M-68DS diluent (Shenzhen Meyer biomedical electronics, Inc.) to form a first sample. The first sample solution is sent to a DC impedance detector, an impedance electrical signal is detected, and a red blood cell region (particles included therein are denoted as a particle group 1, see fig. 11) is distinguished according to a threshold value preset by the system, to obtain a first histogram.

In the classification module, 20 μ L of anticoagulated whole blood sample taken from the same blood sample was mixed with 1mL of M-68LD Lyse and 20 μ LM-68FD dyes (both products of Shenzhen Meyer biomedical electronics, Inc.) to Lyse erythrocytes and stain erythrocytes with nucleic acid substances, forming a second reagent. The second sample solution is delivered to an optical detection system where a scattered light detector detects scattered light and a fluorescence detector detects fluorescence. The light intensities of the obtained side scattered light and fluorescence were analyzed, and the white blood cell population was distinguished from other cells (denoted as the population 2), as shown in the scattergram of fig. 12. The forward scattered light intensity of each cell in the leukocyte population is calculated using the aforementioned formula (1) to obtain a derived volume for each cell, thereby obtaining a second histogram.

The first histogram is overlapped with the second histogram, as shown in fig. 13A, wherein the curve of the second histogram below the curve of the first histogram can be clearly seen from the enlarged view of the 100-300fL area (see fig. 13B).

By calculation, the histogram of the particle group 2 is subtracted from the histogram of the particle group 1, obtaining a histogram of a corrected population of red blood cells (see fig. 14).

Example 4: analysis of red blood cells in a sample using NRBC channels

In the CBC module, 4 μ L of anticoagulated whole blood sample taken from one blood sample was mixed with 1.5mL of M-68DS diluent (Shenzhen Meyer biomedical electronics, Inc.) to form a first sample. The first sample solution is sent to a DC impedance detector, an impedance electrical signal is detected, and a red blood cell region (particles included therein are denoted as a particle group 1, see fig. 15) is distinguished according to a threshold value preset by the system, to obtain a first histogram.

In the classification module, 20 μ L of anticoagulated whole blood sample taken from the same blood sample was mixed with 1mL of M-68LD Lyse and 20 μ LM-68FD dyes (both products of Shenzhen Meyer biomedical electronics, Inc.) to Lyse erythrocytes and stain erythrocytes with nucleic acid substances, forming a second reagent. The second sample solution is sent to an optical detection system, scattered light is detected by a scattered light detector, and fluorescence is detected by a fluorescence detector. The obtained forward scattered light and the light intensity of the fluorescence were analyzed to distinguish the particle group of white blood cells from other cells (denoted as particle group 2), as shown in the scattergram of fig. 16. The forward scattered light intensity of each cell in the leukocyte population is calculated using the aforementioned formula (1) to obtain a derived volume for each cell, thereby obtaining a second histogram.

The first histogram is overlapped with the second histogram, as shown in FIG. 17A, wherein the curve of the second histogram below the curve of the first histogram can be clearly seen from the enlarged view of the 150-300fL region (see FIG. 17B).

By calculation, the histogram of the particle group 2 is subtracted from the histogram of the particle group 1, obtaining a histogram of a corrected population of red blood cells (see fig. 18).

Example 5: analysis of red blood cells in a sample using electrical impedance

In the CBC module, 4 μ L of anticoagulated whole blood sample was mixed with 1.5mL of M-68DS diluent (Shenzhen Meyer biomedical electronics, Inc.) to form a first sample solution. The first sample solution is sent to a DC impedance detector, an impedance electrical signal is detected, and a first histogram (in which the particles included are denoted as a particle group 1, see fig. 19) is obtained by analysis.

Also in the CBC module, a 4 μ L sample of anticoagulated whole blood from the same sample is mixed with 1mL of M-68LD Lyse to Lyse the red blood cells to form a second reagent solution. The M-68LD Lyse is an aqueous solution containing a cationic surfactant, a nonionic surfactant and an anionic compound. The second sample is sent to a DC impedance detector, the impedance electrical signal is detected, and a second histogram is obtained by analysis (the particles contained therein are denoted as particle group 2, see fig. 20).

The first histogram is overlapped with the second histogram, as shown in fig. 21A, wherein the curve of the second histogram below the curve of the first histogram can be clearly seen from the enlarged view of the 100-200fL area (see fig. 21B).

By calculation, the histogram of the particle group 2 is subtracted from the histogram of the particle group 1, obtaining a histogram of a corrected population of red blood cells (see fig. 22).

The second sample solution contains a hemolytic agent, and thus the cells shrink to some extent and change in volume. This results in a non-uniform leukocyte volume of the population 2 through the leukocyte volumes in the population 1. It was found that the tendency of the cell volume change is related to the hemolytic agent used, and that the volume adjustment factor K can be obtained statistically, which is used for the calculation of the derived volume, thus making the volume distribution of leukocytes comparable in the first and second histograms. That is, the white blood cells in the particle group 2 have a functional mapping relationship with the white blood cell volume of the particle group 1, and VOL2 is K × VOL 1.

In order to obtain the above coefficient K, detection may be performed using a sample containing only leukocytes as the first test sample and the second test sample, respectively. The volumes corresponding to the peaks of the two histograms are compared, or the mean cell volume of the two is calculated, obtaining the value of K.

Comparative example 1

The same blood sample as in example 1 was taken, and added to a plethysmometer for low-speed centrifugation to remove the leukocyte layer and the plasma layer. The centrifuged sample is then replenished with diluent to the initial volume. Histograms of the red blood cell populations directly obtained were measured by an electrical impedance method on the same apparatus as in example 1, as shown in fig. 23.

By comparison, the red blood cell counts and related parameters obtained in example 1 and comparative example 1 are shown in Table 1 below, wherein the white blood cell content of the sample was determined to be 291.6X 10^ 9/L. As can be seen from Table 1, the red blood cell counts and morphological parameters obtained by the two methods are consistent with each other.

TABLE 1 Red blood cell count and associated parameter values

The foregoing is merely illustrative of and illustrative of exemplary embodiments and examples of the present invention and is not intended to limit the scope of the invention accordingly. Under the inventive concept of the present invention, the equivalent structural changes made by the contents of the present specification and the attached drawings, or the direct/indirect application to other related technical fields, are all included in the scope of the present invention.

Claims

A method of analyzing red blood cells in a blood sample, the method comprising the steps of:

obtaining a first sample from the blood sample, and mixing the first sample with a first reaction reagent to obtain a first suspension, wherein in the first suspension, red blood cells basically maintain the original shape;

obtaining a second sample from said blood sample, and mixing said second sample with a second reagent comprising a hemolysing agent to obtain a second suspension, wherein red blood cells are lysed in said second suspension;

detecting the first suspension to obtain a first electric signal of each cell, and obtaining first integral distribution information of the cells according to the first electric signal;

detecting said second suspension to obtain cell-by-cell optical information, identifying leukocytes from said optical information, obtaining second volumetric distribution information of leukocytes from at least forward scattered light information of said leukocytes, or

Detecting the second suspension to obtain a second electric signal of each cell, and obtaining third volume distribution information of white blood cells according to the second electric signal;

and correcting the first volume distribution information by using the second volume distribution information or the third volume distribution information to obtain corrected volume distribution information of the red blood cells in the blood sample.
The method of claim 1, wherein the optical information comprises forward scattered light intensity.
The method of claim 2, wherein the optical information further comprises a side-scattered light intensity selected from a medium angle scattered light intensity and/or a high angle scattered light intensity.
The method of claim 3, wherein the method further comprises: and distinguishing basophil subpopulations according to the optical information.
The method of claim 1, wherein said second reagent further comprises a fluorescent dye, or said second sample is further mixed with a third reagent comprising a fluorescent dye to obtain said second suspension.
The method of claim 5, wherein the optical information further comprises fluorescence intensity.
The method of claim 6, wherein the method further comprises: and distinguishing the nucleated red blood cell and the leukocyte particle group according to the optical information.
The method of claim 6, wherein the method further comprises: and distinguishing basophil subpopulations according to the optical information.
The method of claim 5, wherein the optical information includes side scatter light intensity and fluorescence intensity.
The method of claim 9, wherein the method further comprises: differentiating the subpopulations of leukocytes into subpopulations of lymphocytes, monocytes, neutrophils, and eosinophils based on the optical information.
The method of claim 1, wherein the method further comprises: differentiating the leukocyte subpopulations into lymphocyte, monocyte and granulocyte subpopulations based on the second electrical signal.
The method of claim 1, wherein the first, second and third volumetric distribution information and corrected volumetric distribution information are histograms.
The method according to claim 1, wherein the method further comprises outputting at least one information selected from the group consisting of: red blood cell count, mean red blood cell volume (MCV), mean red blood cell hemoglobin content (MCH), mean red blood cell hemoglobin concentration (MCHC), red blood cell distribution width coefficient of variation (RDW-CV), red blood cell distribution width standard deviation (RDW-SD), and packed red blood cell volume (HCT); and/or obtaining a total white blood cell count and a count of a subpopulation of white blood cells from the white blood cell distribution information.
The method of claim 1, wherein the method further comprises outputting the corrected volume distribution information, and optionally first and second volume distribution information, or optionally first and third volume distribution information.
The method according to claim 13 or 14, wherein the outputted information is marked when the white blood cell count exceeds a first threshold value, or when the difference in red blood cell count before and after correction exceeds a second threshold value.
A blood analysis system comprising a sampling part, a reagent supplying part, a reaction part, a detection part, a controller, and a processor, wherein:

the sampling part is configured to obtain at least two samples from a blood sample and convey the samples to the reaction part respectively;

the reagent supply part configured to supply a required reaction reagent to the reaction part;

the reaction part comprises at least one reaction chamber, is used for mixing a first sample with a first reaction reagent to prepare a first suspension, and is used for mixing a second sample with a second reaction reagent to prepare a second suspension, and the reaction part is configured to supply the first suspension and the second suspension to the detection part respectively;

the detection part comprises an electrical detection part and an optical detection part, wherein the electrical detection part is configured to enable the particles in the first suspension to pass through the resistance detector one by one to obtain a first electric signal of the particles in the first suspension, and the optical detection part is configured to enable the particles in the second suspension to pass through the optical detector one by one to obtain optical information of the particles in the second suspension;

the controller is coupled with the sampling part, the reagent supply part, the reaction part and the detection part and controls the actions of the sampling part, the reagent supply part, the reaction part and the detection part; and

the processor, the processor and the detection part are coupled, wherein

The processor acquires the electrical signal and the optical information from the detection portion, acquires first volume distribution information from the electrical signal, acquires white blood cell distribution information from the optical information, acquires second volume distribution information using forward scattered light intensity, and corrects the first volume distribution information with the second volume distribution information to acquire corrected volume distribution information of red blood cells in the blood sample.
The blood analysis system of claim 16, wherein the electrical detection portion comprises a pinhole impedance detector or a sheath flow impedance detector.
The blood analysis system of claim 16, wherein the optical detection system comprises a light source, a flow cell allowing passage of cells one by one, a fluid path system, and a scattered light detector.
The blood analysis system of claim 18, wherein the optical information is scattered light intensity, preferably the optical information comprises forward scattered light intensity and side scattered light intensity.
The blood analysis system of claim 18, wherein the processor distinguishes between subpopulations of white blood cells based on the intensity of scattered light, preferably the processor further distinguishes between subpopulations of basophils.
The blood analysis system of claim 18, wherein the optical detection portion further comprises a fluorescence detector and the second reactive reagent further contains a fluorescent dye, or the reagent supply portion is configured to further supply a third reactive reagent containing a fluorescent dye into the reaction chamber of the reaction portion to be further mixed with the second sample to prepare a second suspension.
The blood analysis system of claim 21, wherein the optical information is at least one of scattered light intensity and fluorescence intensity.
The blood analysis system of claim 22, wherein the scattered light intensity comprises a forward scattered light intensity, and the processor distinguishes between a nucleated red blood cell and a leukocyte particle population, preferably a basophil subpopulation, based on the forward scattered light intensity and a fluorescence intensity.
The blood analysis system of claim 22, wherein the scattered light intensity comprises a side scattered light intensity, and the processor distinguishes between lymphocyte, monocyte, neutrophil, and eosinophil subpopulations based on the side scattered light intensity and fluorescence intensity.
A blood analysis system comprising a sampling part, a reagent supplying part, a reaction part, a detection part, and a controller, wherein:

a sampling section configured to take at least two samples from a blood sample and transfer the samples to the reaction section, respectively;

a reagent supply portion configured to supply a required reaction reagent to the reaction portion;

a reaction part including at least one reaction chamber for preparing a first suspension by mixing a first sample with a first reaction reagent and a second suspension by mixing a second sample with a second reaction reagent, and configured to supply the first suspension and the second suspension to the detection part, respectively;

a detection portion comprising an electrical detection portion, wherein the electrical detection portion is configured to pass particles in the suspension one by one through a resistance detector to obtain a first electrical signal of a first suspension and a second electrical signal of a second suspension;

a controller coupled to the sampling part, the reagent supplying part, the reaction part and the detection part, for controlling the operations of the sampling part, the reagent supplying part, the reaction part and the detection part; and

a processor coupled with the detection portion, wherein

The processor obtains the first and second electrical signals from the detection portion, obtains first volume distribution information from the first electrical signal, obtains third volume distribution information from the second electrical signal, and corrects the first volume distribution information with the third volume distribution information to obtain corrected volume distribution information of red blood cells of the blood sample.
The blood analysis system of claim 16 or 25, wherein the blood analysis system further comprises an output device.
The blood analysis system according to claim 26, wherein the volume distribution information is a histogram, preferably the output device is configured to output the corrected histogram, and optionally the first and second histograms or the first and third histograms.
The blood analysis system of claim 26, wherein the processor obtains at least one parameter selected from the group consisting of: red blood cell count, mean red blood cell volume (MCV), mean red blood cell hemoglobin content (MCH), mean red blood cell hemoglobin concentration (MCHC), red blood cell distribution width coefficient of variation (RDW-CV), red blood cell distribution width standard deviation (RDW-SD), and packed red blood cell volume (HCT); and/or obtaining a total count of white blood cells and a count of lymphocyte, monocyte and granulocyte subpopulations of white blood cells from the second electrical signal; preferably, the output device is configured to output the parameter.
The blood analysis system of claim 27 or 28, wherein the processor compares a white blood cell count with a first threshold value, and when the white blood cell count is greater than the first threshold value, or compares a difference in red blood cell counts before and after correction with a second threshold value, and when the difference is greater than the second threshold value, flags the outputted red blood cell-related information.
A computer-readable storage medium, on which a computer program is stored, which computer program, when being executed by a processor, carries out the steps of:

acquiring a first electric signal obtained by detecting a first suspension obtained by mixing a first sample acquired from a blood sample with a first reaction reagent, and obtaining first integral distribution information from the first electric signal, wherein the electric signal is obtained by detecting the first suspension by an electric detecting portion;

acquiring optical information obtained by detecting a second suspension obtained by mixing a second sample obtained from the blood sample with a second reaction reagent containing at least a hemolytic agent, and obtaining leukocyte distribution information from the optical information, and obtaining second volumetric distribution information using a forward scattered light intensity,

or

Acquiring a second electric signal obtained by detecting a second suspension obtained by mixing a second sample obtained from a blood sample with a second reaction reagent containing at least a hemolytic agent, and obtaining third volume distribution information from the second electric signal, the electric signal being obtained by detecting the second suspension by the electric detection portion; and

and correcting the first volume distribution information by using the second volume distribution information or the third volume distribution information to obtain corrected volume distribution information of the red blood cells in the blood sample.