CN105986003B - White blood cell counting method and device and cell analyzer - Google Patents
White blood cell counting method and device and cell analyzer Download PDFInfo
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- CN105986003B CN105986003B CN201510075567.XA CN201510075567A CN105986003B CN 105986003 B CN105986003 B CN 105986003B CN 201510075567 A CN201510075567 A CN 201510075567A CN 105986003 B CN105986003 B CN 105986003B
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Abstract
A method and apparatus for counting leukocytes is provided, which counts an initial count value of leukocytes from a second leukocyte measurement system of a cell analyzer, and corrects the initial count value using the number of lipid particles acquired from a first leukocyte measurement system, thereby reducing interference of lipid particles in the second leukocyte measurement system with the leukocyte count and accurately counting leukocytes. According to the white blood cell counting method and the white blood cell counting device, a cell analyzer is further provided.
Description
Technical Field
The invention relates to the field of cell analyzers, in particular to a method and a device for counting white blood cells and a cell analyzer.
Background
A blood cell analyzer is an instrument that can detect cells in blood, and can count and classify cells such as White Blood Cells (WBCs), red blood cells, platelets, nucleated red blood cells, reticulocytes, and the like.
The most common method for realizing the white blood cell detection by the blood cell analyzer is a laser scattering method, wherein cells flowing through a detection area are irradiated by laser, and light signals reflected or scattered by the cells are collected to classify and count the white blood cells. The optical signals include forward scattered light, side scattered light, fluorescence, and the like. The forward scattering light signal reflects the size information of the cell, the side scattering light signal reflects the complexity of the internal structure of the cell, and the fluorescence signal reflects the content of substances such as DNA and RNA which can be stained by fluorescent dye in the cell. The scattered light signals can be used to classify the white blood cells and obtain the count value of the white blood cells.
The common five-classification cell analyzer comprises a first leukocyte measurement system and a second leukocyte measurement system, wherein reaction reagents, scattered light to be collected and subsequent data analysis and processing adopted by the first leukocyte measurement system and the second leukocyte measurement system after sample classification are different, the first leukocyte measurement system is generally also called as a DIFF channel or a 4DIFF channel, the first leukocyte measurement system adopts three optical signals of forward scattered light, lateral scattered light, fluorescence and the like to classify and count cells, and leukocytes can be classified into four types of cells such as lymphocyte population, monocyte population, eosinophil population, neutrophils, basophil population and the like; a second leukocyte measurement system, also commonly referred to as BASO channel, can separate leukocytes into basophils and other leukocytes, and can obtain a leukocyte count value and a basophil count value. Five classifications of white blood cells and white blood cell counts can be achieved by the first white blood cell measurement system and the second white blood cell measurement system.
In practical work, since the BASO channel has the advantages of simplicity, convenience, rapidness, low cost and high accuracy, the reliability of the BASO channel is generally accepted clinically, and the BASO channel is often used as the leukocyte count in an output report. However, when lipid particles are present in a blood sample, such as after fat milk injection, white blood cell counting using BASO channels may result in a false increase in white blood cell count, which may affect clinical judgment. Therefore, how to eliminate the influence of lipid particles on the white blood cell count of BASO channels becomes a problem to be solved urgently.
Disclosure of Invention
The application provides a method and a device for counting white blood cells and a cell analyzer, and provides a new scheme for obtaining the white blood cell value from a second white blood cell measuring system of the cell analyzer.
According to a first aspect, there is provided in one embodiment a method of white blood cell counting, comprising:
counting an initial white blood cell count value from a second white blood cell measurement system of the cell analyzer, wherein the second white blood cell measurement system is a measurement system that can classify white blood cells into at least basophils and other white blood cells; acquiring the number of lipid particles from a first leukocyte measurement system of a cell analyzer, wherein the first leukocyte measurement system is a measurement system capable of classifying leukocytes into at least four types of cells, namely lymphocytes, monocytes, eosinophils, neutrophils, and basophils; the initial white blood cell count value is corrected by the number of lipid particles to obtain a corrected white blood cell count value.
According to a second aspect, there is provided in an embodiment a white blood cell counting device comprising: an initial counting module, a lipid particle number obtaining module and a correction module, wherein,
the initial counting module is used for counting the initial counting value of the white blood cells from a second white blood cell measuring system of the cell analyzer, wherein the second white blood cell measuring system is a measuring system which can at least classify the white blood cells into basophils and other white blood cells; the lipid particle number acquisition module is used for acquiring the lipid particle number from a first leukocyte measurement system of the cell analyzer, wherein the first leukocyte measurement system is a measurement system capable of classifying leukocytes into at least four types of cells, namely lymphocyte population, monocyte population, eosinophil population, neutrophil and basophil population; and the correction module is used for correcting the initial counting value of the white blood cells by using the lipid particle number to obtain a corrected counting value of the white blood cells.
According to a third aspect, there is provided in one embodiment a cellular analyzer comprising: the device comprises an optical detection device, a conveying device and the white blood cell counting device, wherein the conveying device is used for conveying the sample liquid to be detected into the optical detection device; the optical detection device is used for irradiating the detected sample liquid flowing through the detection area, collecting various optical signals generated by the cells due to the irradiation of the light, and converting the optical signals into corresponding electric signals for output; the white blood cell counting device is used for receiving and processing the electric signal output by the optical detection equipment.
According to the method and the device for counting white blood cells of the embodiment, the white blood cell number value of the second white blood cell measuring system is corrected through the lipid particle number obtained from the first white blood cell measuring system, so that when the white blood cells are counted from the second white blood cell measuring system, the interference caused by the lipid particles can be reduced, and the white blood cell counting can be realized more accurately.
Drawings
FIG. 1a is a schematic diagram of a blood cell analyzer according to an embodiment;
FIG. 1b is a block diagram of a white blood cell counting device according to an embodiment of the present disclosure;
FIG. 1c is a block diagram of a white blood cell counting device according to another embodiment of the present disclosure;
fig. 1d is a block diagram of an identification module according to an embodiment of the present disclosure;
FIG. 2 is a flow chart of a method for counting leukocytes according to an embodiment of the present disclosure;
FIG. 3 is a flowchart illustrating a method for counting white blood cells according to a first initial white blood cell count value according to an embodiment of the present disclosure;
FIG. 4 is a side scatter-forward scatter plot of a lipid particle sample in a second white blood cell measurement system according to an embodiment of the present disclosure;
FIG. 5a is a side scatter-forward scatter plot of a lipid particle sample in a first white blood cell measurement system according to an embodiment of the present disclosure;
FIG. 5b is a schematic view of FIG. 5a with other particles removed;
FIG. 5c is a three-dimensional scattergram of a lipid particle sample in a first white blood cell measurement system according to an embodiment of the present disclosure;
FIG. 6 shows a WBCB' comparison to WBC reference;
FIG. 7 is a flowchart illustrating a method for counting white blood cells according to a second initial white blood cell count value according to an embodiment of the present application;
fig. 8 is a schematic diagram illustrating a scatter plot of a second white blood cell measurement system in a visualized manner according to an embodiment of the present application;
FIG. 9 is a flow chart of identifying lipid particles based on sub-regions according to an embodiment of the present application;
fig. 10a, 10b and 10c are schematic diagrams of dividing sub-regions according to an embodiment of the present application.
Detailed Description
Referring to fig. 1a, a schematic diagram of a blood cell analyzer is shown, the blood cell analyzer includes an optical detection device 20, a conveying device 30, a data processing device 40 and a display device 50.
The transport device 30 is used for transporting the sample liquid (e.g. the blood sample to be tested) reacted with the reagent to the optical detection device 20. The transport device 30 typically comprises transport lines and control valves through which the sample fluid is transported into the optical detection device 20.
The optical detection device 20 is used for irradiating the sample liquid flowing through the detection area thereof with light, collecting various optical signals (such as scattered light signals and/or fluorescent signals) generated by the cells due to the light irradiation, and converting the optical signals and the corresponding electrical signals into corresponding electrical signals, wherein the information carried by the optical signals and the corresponding electrical signals corresponds to the characteristics of the cell particles, and can be used as cell particle characteristic data, the forward scattered light signals reflect the size information of the cells, the side scattered light signals reflect the complexity of the internal structure of the cells, and the fluorescent signals reflect the content of substances, such as DNA and RNA, which can be stained by fluorescent dyes in the cells. In the embodiment shown in FIG. 1a, optical inspection apparatus 20 may include a light source 1025, a flow cell 1022 as the inspection region, a forward scatter signal collector 1023 disposed on the optical axis, a side scatter signal collector 1026 disposed to the side of the optical axis, and a fluorescence signal collector 1027.
The blood sample is divided according to the need, the sample liquid of the first white blood cell measuring system and the second white blood cell measuring system after reacting with different reagents passes through a flowing chamber 1022 for providing a detection area under the wrapping of sheath liquid, the light beam emitted by a light source 1025 irradiates the detection area 1021, each cell particle in the sample liquid emits scattered light after being irradiated by the light beam, a light collecting device collects the scattered light, for example, when the sample liquid of the DIFF channel passes through the detection region 1021, the forward scattered light signal collection means 1023, the side scattered light signal collection means 1026 and the fluorescence signal collection means 1027 collect forward scattered light, side scattered light and fluorescence signals respectively, forward scattered light signal collection apparatus 1023 and side scattered light signal collection apparatus 1026 collect forward scattered light and side scattered light signals, respectively, as the BASO channel sample fluid passes through detection region 1021. The collected and shaped light is irradiated to the photoelectric sensor, and the photoelectric sensor converts the light signal into a corresponding electrical signal and outputs the electrical signal to the data processing device 40.
The data processing device 40 performs analysis processing on the received particle characteristic data to realize analysis of the blood sample to be tested. In the present embodiment, the data processing device 40 includes a white blood cell counting device for receiving the electrical signal representing the optical signal output from the optical detection device, generating a scattergram from the characteristic data set of each particle by using the optical signal of each particle as a characteristic data set representing the particle. In the embodiment of the present invention, the scattergram is a set consisting of characteristic data sets of each cell particle, and may be stored in a storage device in a digitized form or may be visually presented on a display interface. The white blood cell counting device further processes the particle scatter plot to obtain a five-point classification of white blood cells and a particle count. In the present embodiment, the first leukocyte measurement system of the leukocyte counting device generates a scattergram from the optical signal of the sample collected by the light collection device, and obtains the results of counting the four types of cells, i.e., the lymphocyte population, the monocyte population, the eosinophil population, the neutrophil population, and the basophil population, from the scattergram and a predetermined threshold value or gating method, and the second leukocyte measurement system of the leukocyte counting device obtains a scattergram from the characteristic data group of each cell particle from the scattered light signal of the sample collected by the light collection device, and obtains the leukocyte count value and the basophil count value from the predetermined threshold value or gating method. The white blood cell counting device also acquires a lipid particle counting value from the first white blood cell measuring system, and eliminates the counting interference of the lipid particles on the white blood cells in the second white blood cell measuring system through the lipid particle counting value, so as to correct the white blood cell counting value obtained in the second white blood cell measuring system.
The display device 50 is electrically coupled to the data processing apparatus 40 and is used for displaying the analysis result output by the data processing apparatus 40, wherein the analysis result can be a graph, a text description and/or a table. In this embodiment, the display device 50 may output various visualized scatter diagrams and/or various cell classification results.
Referring to fig. 1b, a block diagram of a white blood cell counting device disclosed in this embodiment is shown, the white blood cell counting device includes: the device comprises an initial counting module 1, a lipid particle number acquisition module 2 and a correction module 3. Wherein the content of the first and second substances,
the initial counting module 1 is used for counting the initial counting value of the white blood cells from a second white blood cell measuring system of the cell analyzer, wherein the second white blood cell measuring system is a measuring system capable of classifying the white blood cells into basophils and other white blood cells. For example, the initial white blood cell count value is counted based on a scatter chart obtained by the second white blood cell measurement system. In one embodiment, the initial counting module 1 comprises a first initial counting unit; in another embodiment, the initial counting module 1 comprises a second initial counting unit. The first initial counting unit is used for generating a scatter diagram of a second white blood cell measuring system according to scattered light signals of a measured blood sample collected by the second white blood cell measuring system of the cell analyzer, and counting the number of cells in a non-blood shadow area in the scatter diagram of the second white blood cell measuring system to obtain a first initial white blood cell count value; the second initial counting unit is used for generating a scatter diagram of the second white blood cell measuring system according to scattered light signals of the measured blood sample collected by the second white blood cell measuring system of the cell analyzer, and counting the number of particles in a white blood cell area in the scatter diagram of the second white blood cell measuring system to obtain a second initial white blood cell count value. In a specific embodiment, the scattered light signals collected by the second white blood cell measurement system include forward scattered light signals and side scattered light signals, and the number of particles in the non-ghost region or the number of particles in the white blood cell region is counted based on a two-dimensional scattergram of the forward scattered light and the side scattered light signals.
The lipid particle count acquisition module 2 is configured to acquire the lipid particle count from a first leukocyte measurement system of the cell analyzer, wherein the first leukocyte measurement system is a measurement system capable of classifying leukocytes into at least four types of cells, i.e., a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil population, and a basophil population. In a specific embodiment, the lipid particle count acquisition module 2 includes a lipid particle counting unit configured to generate a scatter diagram according to an optical signal of a blood sample to be measured collected by a first white blood cell measurement system of the cell analyzer, identify lipid particles based on the scatter diagram, and acquire a first lipid particle count value or a second lipid particle count value, wherein the first lipid particle count value is the number of lipid particles in a non-hemogram region, and the second lipid particle count value is the number of lipid particles in a white blood cell region. For example, the light signals collected by the first leukocyte measurement system include forward scattered light signals and side scattered light signals, and on the two-dimensional scattergram of the forward scattered light signals and the side scattered light signals, lipid particles can be identified well distinguished from other particles, and the first lipid particle count value is obtained by counting the number of lipid particles in a non-ghost region or the second lipid particle count value is obtained by counting the number of lipid particles in a leukocyte region. Preferably, the optical signals collected by the first leukocyte measurement system further include fluorescence signals, and in a three-dimensional scattergram of forward scattered light, side scattered light and fluorescence signals, the lipid particles are more clearly distinguished from other particles and can be better identified, and the obtained first lipid particle count value and the second lipid particle count value are more accurate.
The correction module 3 is used for correcting the initial white blood cell count value by using the lipid particle number to obtain a corrected white blood cell count value. In one embodiment, the modification module 3 is configured to subtract the first lipid particle count value from the first initial white blood cell count value to obtain a modified white blood cell count value; in another embodiment, the modification module 3 is configured to subtract the second lipid particle count value from the second initial white blood cell count value to obtain a modified white blood cell count value.
In a preferred embodiment, the white blood cell counting device may further comprise a labeling module 6 for labeling the lipid particles differently from the labeled white blood cells in the scatter plot of the second white blood cell measurement system. In general, the scatter plot may be presented in a visual form on a display interface of a display device.
Referring to fig. 1c, in another preferred embodiment, the white blood cell counting apparatus may further include an identification module 5, the identification module 5 is used for identifying lipid particles of a scatter diagram of the second white blood cell measurement system, and in a specific embodiment, the identification module 5 may map the lipid particles in the second white blood cell measurement system by the number of lipid particles in the first white blood cell measurement system. Specifically, a scatter diagram of the first leukocyte measurement system and a scatter diagram of the second leukocyte measurement system may be divided into a plurality of corresponding sub-regions, the number of lipid particles in each sub-region in the first leukocyte measurement system is counted, and then the number of particles in the corresponding sub-region in the second leukocyte measurement system is selected as lipid particles through a certain functional relationship. Further, the labeling module 6 labels the lipid particles identified by the identification module 5 on the scatter diagram of the second white blood cell measurement system in a manner different from that of the labeled white blood cells, and presents the labeled white blood cells to the user in a visual form.
Referring to fig. 1d, the identification module 5 includes: a first area dividing unit 51, a second area dividing unit 52, a counting unit 53, and an identifying unit 54, wherein,
the first region dividing unit 51 is configured to divide a scatter diagram of the first white blood cell measurement system into a plurality of first sub-regions along a direction of laterally scattering light; the second region dividing unit 52 is configured to divide the scattergram of the second white blood cell measurement system into a plurality of second sub-regions along the direction of the laterally scattered light. The first region dividing unit 51 and the second region dividing unit 52 divide the sub-regions in the same manner, so that each first sub-region and each second sub-region have a one-to-one correspondence relationship. It should be noted that, in this embodiment, the dividing sequence of the first area dividing unit 51 and the second area dividing unit 52 is not limited.
The counting unit 53 is configured to count the number of lipid particles in the first sub-region.
The identification unit 54 is configured to select particles in the second sub-region and identify the particles as lipid particles, so that the number of lipid particles in the first sub-region is the same as the number of lipid particles in the second sub-region corresponding to the first sub-region; alternatively, the ratio of the number of lipid particles in each first subregion to the number of lipid particles in the corresponding second subregion is the same.
Example 1:
based on the above white blood cell counting device, the present embodiment also discloses a white blood cell counting method, please refer to fig. 2, which is a flow chart of the white blood cell counting method, and the method includes the following steps:
step S100, initial counting. Counting an initial white blood cell count value from a second white blood cell measurement system of the cell analyzer, wherein the second white blood cell measurement system is a measurement system capable of classifying at least white blood cells into basophils and other white blood cells, and collecting forward scattered light signals and side scattered light signals of the cells flowing through the detection area to form a scatter diagram. In a specific embodiment, the initial count value may be the number of particles in the non-ghost region in the second white blood cell measurement system scatter plot; in another embodiment, the initial count value may also be the number of particles in the white blood cell region in the scatter plot of the second white blood cell measurement system. The ghost is a cell fragment structure formed after a blood sample is subjected to sample pretreatment, cells such as red blood cells or platelets and the like in the blood sample are subjected to low permeation or reagent treatment, cell membranes are broken, particle size is small, forward scattered light signals are small, and therefore, a threshold value can be set for the forward scattered light signals of the ghost according to experience, so that a distribution area of the ghost can be defined in a side scattered light-forward scattered light scatter diagram. The region other than the distribution region of the ghost is referred to as a non-ghost region. The data of the scattered light signals of a particle can be compared with respective thresholds to determine whether the particle belongs to a shadow region or a non-shadow region. Also, empirically, it is possible to determine the white blood cell region in the side scatter-forward scatter diagram by setting a threshold and gating, and to determine whether a particle belongs to the white blood cell region by comparing the data of the scattered light signal of the particle with the respective threshold or gating.
Step S200, lipid particle count acquisition. The method comprises the steps of identifying lipid particles from a first leukocyte measurement system of a cell analyzer, and acquiring a first lipid particle count value or a second lipid particle count value, wherein the first leukocyte measurement system is a measurement system capable of classifying leukocytes into at least four types of cells including lymphocyte groups, monocyte groups, eosinophil groups, neutrophils and basophil groups, and collecting forward scattered light signals and side scattered light signals flowing through cells in a detection area to form a scatter diagram, or forward scattered light information, side scattered light signals and fluorescence signals to form a three-dimensional scatter diagram. In a specific embodiment, the first lipid particle count value is the number of lipid particles in the non-haemographic region and the second lipid particle count value is the number of lipid particles in the leukocyte region. The non-ghost area of the first measurement system scattergram is an area on the scattergram on which the non-ghost area of the second measurement system scattergram of the side scattered light signal and the forward scattered light signal is mapped, and the leukocyte area of the first measurement system scattergram is an area on the scattergram on which the leukocyte area of the second measurement system scattergram is mapped, that is, the non-ghost area and the leukocyte area of the first measurement system scattergram have the same threshold value or threshold value as the non-ghost area and the leukocyte area of the second measurement system scattergram, respectively.
It should be noted that the execution sequence of step S100 and step S200 may be determined based on the operation sequence of the second white blood cell measurement system and the first white blood cell measurement system, and of course, the processing of step S100 and step S200 may be started after all data are collected.
And step S300, correction. The initial white blood cell count value is corrected by the number of lipid particles to obtain a corrected white blood cell count value. In a specific embodiment, the value of the lipid particles in the non-ghost or white blood cell region can be subtracted from the initial count value of the region to obtain the white blood cell count value of the region.
The correction of the cell count is specifically described below.
In one embodiment, referring to fig. 3, a flowchart of a white blood cell counting method according to the present embodiment is shown when performing a correction according to a first initial white blood cell count value, the method includes the following steps:
step S110, a first initial count. And generating a scatter diagram of the second leukocyte measurement system according to scattered light signals of the measured blood sample collected by the second leukocyte measurement system of the cell analyzer, and counting the number of cells in a non-hemogram area in the scatter diagram of the second leukocyte measurement system to obtain a first initial leukocyte count value. In a specific embodiment, the scattered light collected by the second white blood cell measurement system includes forward scattered light signals and side scattered light signals, and the number of cells in the non-ghost area is counted based on a two-dimensional scattergram of the forward scattered light signals and the side scattered light signals.
Referring to fig. 4, a side scattered light-forward scattered light scattergram of an example of a sample containing lipid particles in a second white blood cell measurement system is shown. The non-shadow areas are located above H1-H1 and the shadow areas are located below H1-H1, although in other embodiments, other areas, such as a closed area, may be present in the non-shadow areas. In the non-ghost area, the a area is the area where white blood cells are distributed, and is defined as the area of white blood cells, i.e. the area of particle distribution shown by the white blood cell population, such as the a area distributed with white blood cell subsets such as lymphocytes, monocytes, neutrophils, eosinophils, basophils, etc. Z shows lipid particles which in the scatter plot exhibit the characteristics of an S-curve, with the low end of the signal passing through the leukocyte clusters, thus affecting the leukocyte count. In this example, the first initial white blood cell count value is the sum of the number of white blood cells and lipid particles in the non-ghost region.
It should be noted that in other embodiments, the extent of the non-shadow region may be set in the system, such as increasing or decreasing the extent of the non-shadow region. Alternatively, the areas other than the angiogram areas on the scattergram may be defined as non-angiogram areas by other methods, for example, by adjusting gating rules of the angiogram areas according to the detection system to find the angiogram areas.
Step S210, the first lipid particle count. Since the fluorescence intensity of the lipid particle is lower than that of the white blood cell (including lymphocyte, monocyte, neutrophil, basophil and eosinophil), the lipid particle can be significantly distinguished from the white blood cell on the scatter diagram of the fluorescence signal, so that the count value of the lipid particle can be identified and obtained from the two-dimensional or three-dimensional scatter diagram of the first white blood cell measurement system including fluorescence and scattered light signals. In a specific embodiment, the first white blood cell measurement system collects scattered light signals of a blood sample to be measured, and generates a side scattered light-forward scattered light scatter diagram, please refer to fig. 5a, white blood cell population and lipid particles can be distinguished, lipid particles can be identified, and the number of particles in a non-ghost area can be counted as a first lipid particle count value; of course, the figure can also be treatedParticles other than the lipid particle were physically removed to obtain a lipid particle scattergram, and as shown in fig. 5b, the number of particles in the non-ghost region was counted as a first lipid particle count value. Preferably, the fluorescence signal is also collected to generate a three-dimensional scattergram, and in this embodiment, the three-dimensional scattergram is generated according to the forward scattered light, the side scattered light and the fluorescence of the blood sample to be measured collected by the first white blood cell measurement system of the cell analyzer, and on the three-dimensional scattergram, the differentiation between the lipid particle and other particles is more obvious, and the identification of the lipid particle is more accurate. Referring to FIG. 5c, a three-dimensional scattergram of a sample containing lipid particles in a first white blood cell measurement system is shown. In fig. 5c, the region a is a white blood cell distribution region, and the region Z is a lipid particle distribution region, and as can be seen from fig. 5c, in the three-dimensional scattergram of the first white blood cell measurement system, the distribution regions of white blood cells and lipid particles can be distinguished significantly without overlapping, the lipid particles (region Z) can be classified by the three-dimensional scattergram of the first white blood cell measurement system, and the first lipid particle count value WBC can be obtained by counting the number of particles of the lipid particles located in the non-ghost region (which can be mapped from the non-ghost region of the second white blood cell measurement system)DL. Of course, the map may be processed to remove particles other than the lipid particle to obtain a lipid particle scattergram, and the number of particles in the non-ghost area may be counted as the first lipid particle count value.
In step S310, the white blood cell count value is corrected. The first initial white blood cell count value WBCBSubtract the non-ghost region lipid particle count value WBC in the second leukocyte measurement SystemBLObtaining the white blood cell count value WBC of the corrected non-shadow areaB'。
Although not wishing to be bound by theory, it was found in the study that since lipid particles are not cells, the population of particles appeared similarly in position on the respective scattergrams, i.e. the distribution areas were uniform or functionally related, when detected using the same scattered light signal. Although the first and second leukocyte measurement systems use different reagents, since both the first and second leukocyte measurement systems use forward scattered light and side scattered light signals for cell measurement, the distribution regions of the lipid particles in the two systems are consistent or have a functional relationship. Please refer to fig. 5a, which is a side scattered light-forward scattered light scattergram of the above sample containing lipid particles in the first white blood cell measurement system. Comparing fig. 5a and fig. 4, the lipid particle exhibits similar characteristics of the S-curve in the first and second leukocyte measurement systems.
Since the lipid particles have a uniform or functionally related distribution area in the side scattered light-forward scattered light scattergram of the first and second leukocyte measurement systems, the first lipid particle count value WBC in the first leukocyte measurement system is usedDLThe corresponding count WBC of the non-ghost region lipid particles in the second white blood cell measurement system can be calculatedBLAnd is recorded as:
WBCBL＝f(WBCDL)
in particular embodiments, the function f () may be a multiple, i.e.
WBCBL＝αWBCDL
Where α is a constant, which can be determined from a predetermined or empirical value, in one embodiment α is 1.0, WBC due to the difference between the two leukocyte detection systemsBLAnd WBCDLThe difference is possible, the value range of alpha can be 0.5-2, and the value can be generally 0.8-1.2 to meet the requirement. From this, a corrected white blood cell count value WBC in the second white blood cell measurement system can be obtainedB' is:
WBCB'＝WBCB-WBCBL＝WBCB-αWBCDL
thereby, the white blood cell count value WBC in the second white blood cell measurement system is correctedB'。
For example, the following steps are carried out: in one example, a lipid particle sample is treated by a serum replacement test method to remove lipid particles, and then a white blood cell reference value obtained by a particle counter is 5.96X 109A first lipid particle count value WBC in a first leukocyte measurement systemDL＝2.76×109The non-ghost area lipid particle count value WBC in the second leukocyte measurement system can be obtained by using the coefficient alpha of 1BL＝2.76×109/L, first initial white blood cell count value WBC of non-ghost area in second white blood cell measurement SystemBL＝2.76×109The final white blood cell count value of the second white blood cell measuring system is WBCB'＝5.97×109L, and WBC reference value 5.96X 109the/L correspondence is relatively accurate.
Referring to fig. 6, 237 lipid particle samples were selected, and fig. 6 is a comparison between the white blood cell count value and the standard reference value corrected by the method of the present embodiment, as can be seen from fig. 6, WBCB' the slope of the correlation with the white blood cell count reference (WBC reference) is 1.020, the intercept is-0.038, R2R is a correlation coefficient, indicating that the correlation is good, indicating that the leukocyte count value after correction using the protocol of this example is very close to the true value.
In another embodiment, referring to fig. 7, a flowchart of a white blood cell counting method according to the present embodiment is shown when the correction is performed according to the second initial white blood cell count value, the method includes the following steps:
step S120, second initial counting. And generating a scatter diagram of the second leukocyte measurement system according to scattered light signals of the measured blood sample collected by the second leukocyte measurement system of the cell analyzer, and counting the number of particles in a leukocyte area in the scatter diagram of the second leukocyte measurement system to obtain a second initial leukocyte count value. Unlike the above-described counting of particles in the non-shadowed region, the second initial white blood cell count value in this embodiment is the number of particles in the white blood cell region, which of course also contains a portion of lipid particles, which interferes with the white blood cell count in this region. In this embodiment, the second initial white blood cell count value is the sum of the white blood cell count of the white blood cell region and the lipid particle count in the region.
Step S220, second lipid particle counting. A scatter diagram is generated from scattered light of a blood sample to be measured collected by a first white blood cell measurement system of a cell analyzer, lipid particles are identified based on the scatter diagram of the first white blood cell measurement system, and a second lipid particle count value, which is the number of lipid particles in a white blood cell region, is acquired. The difference from the above embodiment is that the counting range of the lipid particles is different, the number of the lipid particles in the non-ghost region is acquired in step S210, and in the present embodiment, the number of the lipid particles in the leukocyte region is acquired, that is, the counting range of the region may be different. As can be seen from the above description, in the scattergram of the first white blood cell measurement system, white blood cells and lipid particles can be distinguished significantly, and there is no overlap, and lipid particles can be classified by the scattergram of the first white blood cell measurement system. The white blood cell region on the scattergram of the forward scattered light signal and the side scattered light signal of the second white blood cell measurement system is mapped onto the two-dimensional scattergram of the forward scattered light signal and the side scattered light signal of the first white blood cell measurement system, and the second lipid particle count value can be obtained by counting the number of lipid particles located in the white blood cell region. It is also preferred that the lipid particles are better distinguishable from other particles and better identifiable on the three-dimensional scattergram of the forward scattered light, side scattered light and fluorescence signal of the first white blood cell measurement system. And mapping the white blood cell area of the second white blood cell measuring system to a scatter diagram of the forward scattered light signals and the side scattered light signals of the first white blood cell measuring system to obtain the corresponding white blood cell area of the first white blood cell system. After the lipid particles are identified by the three-dimensional scatter diagram, the number of the lipid particles in the corresponding leukocyte region is counted to obtain a second lipid particle count value. In a particular embodiment, the lipid particles are identified and counted in a scatter plot based on the first white blood cell measurement system to obtain a second lipid particle count value, which is also preferably weighted by a function, in particular one function being a multiple function, which may preferably be 1. Of course, in other embodiments, similarly, the two-dimensional or three-dimensional scattergram of the first white blood cell measurement system may be processed to remove particles other than the lipid particle, obtain a lipid particle scattergram, and count the number of particles in the white blood cell region as the second lipid particle count value.
In step S320, the white blood cell region is corrected. The second lipid particle count value is subtracted from the second initial leukocyte count value to obtain a corrected leukocyte count value, which may specifically refer to the calculation manner in step S310, and the principle is similar, and is not described herein again. In a preferred embodiment, when the second lipid particle count value is weighted by the function in step S220, the weighted value is used to correct the white blood cell count value.
In the embodiment, the correction of the white blood cell count value is performed on the white blood cell region, and the pertinence is strong compared with the correction of the white blood cell count value on the whole non-ghost region, so that the result obtained by the correction is more accurate theoretically.
In this embodiment, the advantage that the first white blood cell measurement system can significantly distinguish lipid particles is utilized to count the number of lipid particles in the first white blood cell measurement system, and then the number of lipid particles in the second white blood cell measurement system is calculated according to the characteristic that the distribution of lipid particles of the second white blood cell measurement system and the first white blood cell measurement system has consistency or correlation when the scatter diagram is generated by using the same scattered light detection, so that the influence of the lipid particles on the interference of the counted number of white blood cells in the second white blood cell measurement system is eliminated, and more accurate white blood cell count is obtained.
Example 2:
referring to fig. 2, 3 and 7, the method for counting white blood cells disclosed in this embodiment further includes labeling lipid particles in a scatter diagram of a second white blood cell measurement system, including:
step S400, lipid particle labeling. Referring to fig. 8, in general, the cellular analyzer presents a scatter diagram of the first and second white blood cell measurement systems in a visual form on a display interface of a display device. In this embodiment, the visually displayed scattergram includes at least a two-dimensional scattergram of side scattered light-forward scattered light of the second white blood cell measurement system, and in other embodiments, other scattergrams may be displayed, such as adding dimensions or displaying other scattergrams, according to the user's needs. In order to improve the user experience, the user can visually see the distribution of the lipid particles in the white blood cell area on the scatter diagram, and the lipid particles are marked in a mode different from that of the white blood cells in the visual scatter diagram. In particular embodiments, different symbols and/or different colors may be used for distinction. For example, black plus signs are white blood cell particles and black dots are lipid particles. In other embodiments, different markers may be used to distinguish between white blood cells and lipid particles throughout the scattergram, allowing the user to visually see the distribution of white blood cells and the distribution of lipid particles on the scattergram.
The lipid particles on the scatter diagram of the second leukocyte measurement system are marked in a mode different from a leukocyte marking mode, are displayed on a display interface in a visual mode, and are visually distinguished, so that the user experience is improved, and the research and analysis of the user on the sample are facilitated.
Step S400 and step S320 are not sequentially limited, i.e., the correction of the white blood cell count value and the labeling of the lipid particle on the second white blood cell measurement system may be performed in parallel.
Example 3
The method for counting leukocytes disclosed in this embodiment further includes: referring to fig. 9, a flowchart of identifying lipid particles in a scattergram of a second white blood cell measurement system in this embodiment is specifically illustrated, and includes the following steps:
step S510, dividing the first sub-area. In a scatter diagram of the first white blood cell measurement system, the scatter diagram is divided into a plurality of first sub-regions in the direction of the laterally scattered light. Referring to fig. 10a, 10b and 10c, in an embodiment, the side scattering light direction may be equally divided into N segments, where N is 5 to 20, preferably N is 8, a specific value may be determined according to a resolution of the scattergram, the resolution is high, and N may be larger. In other embodiments, the N segments may not be equally divided.
In step S520, the second sub-area is divided. In a scatter diagram of the second white blood cell measurement system, the scatter diagram is divided into a plurality of second sub-regions in the direction of the laterally scattered light. Referring to fig. 10a, 10b and 10c, in an embodiment, the side scattering light direction may be equally divided into N segments, where N is 5 to 20, preferably N is 8, a specific value may be determined according to a resolution of the scattergram, the resolution is high, and N may be larger. In other embodiments, the N segments may not be equally divided.
In this embodiment, the execution sequence of step S510 and step S520 is not limited. The first sub-regions are divided in the same manner as the second sub-regions, so that each first sub-region has a one-to-one correspondence relationship with each second sub-region, that is, the first sub-regions and the corresponding second sub-regions have the same signal intensity in the direction of the side scattering light. Referring to fig. 10a and 10b, the scatter plots of the first and second white blood cell measurement systems showed consistent signal intensity per unit interval in the direction of side scatter light. In fig. 10a, the scattergram of the first and second white blood cell measurement systems is equally divided into N segments in the lateral scattered light direction, and is not equally divided into N segments in fig. 10b, and each first sub-region corresponds to each second sub-region in the lateral scattered light direction. Referring to fig. 10c, the signal intensities represented by the unit interval in the side scattering light direction in the scattergram of the first white blood cell measurement system and the scattergram of the second white blood cell measurement system are different, specifically: the signal intensity of the scatter diagram of the second white blood cell measurement system in the unit interval in the direction of the side scattered light is smaller than the signal intensity of the scatter diagram of the first white blood cell measurement system in the unit interval in the direction of the side scattered light. Therefore, the range of the scattergram region presented by the second leukocyte measurement system is larger than the range of the scattergram region presented by the first leukocyte measurement system, so that when the first and second subregions are divided in the same dividing manner, the region range of the second subregion corresponding to each first subregion should be larger than the range of the first subregion, and vice versa.
Step S530, counting the first sub-area. The number of lipid particles in the first subregion was counted. As can be seen from the above description, since the lipid particles can be identified in the first white blood cell measurement system, the number of lipid particles in each sub-region can be counted. The number of lipid particles in the ith first subregion is not set to giWherein i is 1 to N. In one embodiment, only the possible memory may be countedG in the first subregion of the lipid particle coinciding with a leukocytei(ii) a In another embodiment, the number of lipid particles in each first subregion can also be counted separately.
Step S540, lipid particle identification. And selecting particles in the second sub-area and identifying the particles as lipid particles, wherein the number of the particles identified as the lipid particles is the same as or has a certain proportional relation with the number of the lipid particles of the corresponding first sub-area. The number m of lipid particles in the second subregioniWherein i is 1 to N, miThe number of lipid particles in the ith second subregion in the scatter plot of the second leukocyte measurement system. In a particular embodiment, m may be calculated using the following formulai：
mi＝T(gi)
The function T () may be a proportional function, such as:
mi＝βgi
in a specific embodiment, the value of β is in a range of 0.5 to 2, and preferably β is 1.0. m isi＝βgiIt was found that the ratio of the number of lipid particles in each first subregion to the number of lipid particles in the corresponding second subregion was the same. The identification of lipid particles for each second sub-region in the second leukocyte measurement system is then completed.
In a specific embodiment, after the scatter plot of the second white blood cell measurement system is divided into several sub-regions, the labeling of the lipid particles may be performed by sub-region. E.g. in finding miThen, m can be randomly selected in the ith second sub-areaiIndividual particles were labeled as lipid particles, thereby completing the labeling of the lipid particles in the scattergram of the second white blood cell measurement system, as shown in fig. 10a, 10b, and 10 c. In other embodiments, other operations may be performed after the lipid particles are identified in the molecular region according to the user's needs.
In this embodiment, by dividing the scatter diagram into several sub-regions, the counting and identification of lipid particles are performed based on the sub-regions, and thus the regional pertinence of the lipid particle identification is further improved. In addition, the lipid particles in the second leukocyte measurement system scatter diagram are marked based on the sub-areas, so that the distribution situation of the lipid particles can be better reflected.
In a further embodiment, the lipid particles identified in the scatter diagram of the first white blood cell measurement system are then determined from the data set of these lipid particles and identified in their distribution positions in the scatter diagram of the second white blood cell measurement system.
Furthermore, after the lipid particles are identified in the scatter diagram of the second white blood cell measurement system, the lipid particles are marked in a different way from the white blood cell marking way and are visually presented to the user.
The technical features or operational steps described in the embodiments of the present invention may be combined in any suitable manner. Those of skill in the art will readily appreciate that the order of the steps or acts in the methods described in connection with the embodiments of the invention may be varied. Accordingly, unless otherwise specified a certain order is required, any order in the drawings or detailed description is for illustrative purposes only and is not necessarily required.
The present invention has been described above with reference to specific examples, but the present invention is not limited to these specific examples. It will be understood by those skilled in the art that various changes, substitutions of equivalents, variations and the like can be made in the embodiments provided by the present invention without departing from the spirit of the invention, and these changes are within the scope of the invention. In addition, the "one embodiment" or "another embodiment" described in many places above are different embodiments, and all or part of them may be combined into one embodiment.
Those skilled in the art will appreciate that all or part of the steps of the various methods in the above embodiments may be implemented by instructions associated with hardware via a program, which may be stored in a computer-readable storage medium, and the storage medium may include: read-only memory, random access memory, magnetic or optical disk, and the like.
The present invention has been described in terms of specific examples, which are provided to aid understanding of the invention and are not intended to be limiting. For a person skilled in the art to which the invention pertains, several simple deductions, modifications or substitutions may be made according to the idea of the invention.
Claims (11)
1. A method of white blood cell counting, comprising:
counting an initial white blood cell count value from a second white blood cell measurement system of the cell analyzer, the second white blood cell measurement system being a measurement system that can classify at least white blood cells into basophils and other white blood cells;
acquiring the number of lipid particles from a first leukocyte measurement system of a cell analyzer, the first leukocyte measurement system being a measurement system that can classify leukocytes into at least four types of cells of a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil population, and a basophil population;
correcting the initial counting value of the white blood cells by using the lipid particle number to obtain a corrected counting value of the white blood cells;
counting initial white blood cell counts from a second white blood cell measurement system of the cellular analyzer includes:
generating a scatter diagram of a second leukocyte measurement system from forward scattered light signals and side scattered light signals of the blood sample under test collected by the second leukocyte measurement system of the cell analyzer;
counting the number of particles in a non-hemogram area in a scatter diagram of a second white blood cell measuring system to obtain a first initial white blood cell count value; or counting the number of particles in the white blood cell area in a scatter diagram of a second white blood cell measuring system to obtain a second initial white blood cell count value;
obtaining the lipid particle count from the first white blood cell measurement system of the cellular analyzer comprises: generating a scatter diagram according to forward scattered light signals and side scattered light signals of a blood sample to be tested, which are collected by a first white blood cell measurement system of a cell analyzer, identifying lipid particles based on the scatter diagram of the first white blood cell measurement system, and acquiring a first lipid particle count value or a second lipid particle count value; the first lipid particle count value is the number of lipid particles in the non-angiographic region; the second lipid particle count value is the number of lipid particles in the leukocyte region;
correcting the initial white blood cell count with lipid particle counts includes: subtracting the first lipid particle count value from the first initial leukocyte count value to obtain a corrected leukocyte count value; or subtracting the second lipid particle count value from the second initial leukocyte count value to obtain a corrected leukocyte count value.
2. The method of claim 1,
the scattered light signals collected by the second white blood cell measurement system comprise forward scattered light signals and side scattered light signals, and the particle number in the non-ghost area or the particle number in the white blood cell area is counted based on the two-dimensional scatter diagram of the forward scattered light signals and the side scattered light signals; and
the light signals collected by the first leukocyte measurement system include at least forward scattered light signals and side scattered light signals, and lipid particles are identified based on a two-dimensional scattergram of the forward scattered light signals and the side scattered light signals.
3. The method of claim 2, wherein the light signals collected by the first white blood cell measurement system further comprise fluorescent signals, and the lipid particle is identified based on a three-dimensional scattergram of forward scattered light signals, side scattered light signals, and fluorescent signals.
4. The method of claim 1 or 2, further comprising:
the lipid particles are labeled in a different way than the labeled leukocytes in the scatter plot of the second leukocyte measurement system.
5. The method of claim 1 or 2, further comprising: identifying lipid particles in the particles detected by the second leukapheresis system:
dividing a scatter diagram of a second white blood cell measuring system into a plurality of second sub-areas along the direction of lateral scattered light;
dividing a scatter diagram of a first white blood cell measuring system into a plurality of first sub-areas along the direction of lateral scattered light;
the dividing mode of the first sub-area is the same as that of the second sub-area, so that each first sub-area and each second sub-area have one-to-one correspondence;
counting the number of lipid particles in the first subregion;
selecting particles in the second sub-area and identifying the particles as lipid particles, so that the number of the lipid particles in the first sub-area is the same as that of the lipid particles in the second sub-area corresponding to the first sub-area; alternatively, the ratio of the number of lipid particles in each first subregion to the number of lipid particles in the corresponding second subregion is the same.
6. A white blood cell counting device, comprising:
an initial counting module for counting an initial count value of white blood cells from a second white blood cell measurement system of the cell analyzer, the second white blood cell measurement system being a measurement system that can classify white blood cells into at least basophils and other white blood cells;
a lipid particle number acquisition module for acquiring a lipid particle number from a first leukocyte measurement system of a cell analyzer, the first leukocyte measurement system being a measurement system capable of classifying leukocytes into at least four types of cells, i.e., a lymphocyte population, a monocyte population, an eosinophil population, a neutrophil, and a basophil;
the correction module is used for correcting the initial counting value of the white blood cells by using the lipid particle number to obtain a corrected counting value of the white blood cells;
the initial counting module comprises:
a first initial counting unit, configured to generate a scatter diagram of a second leukocyte measurement system according to the forward scattered light signal and the side scattered light signal of the blood sample to be measured collected by the second leukocyte measurement system of the cell analyzer, and count the number of cells in a non-shadowed area in the scatter diagram of the second leukocyte measurement system to obtain a first initial leukocyte count value; and/or the presence of a gas in the gas,
a second initial counting unit, configured to generate a scatter diagram of a second leukocyte measurement system according to the forward scattered light signal and the side scattered light signal of the blood sample to be measured collected by a second leukocyte measurement system of the cell analyzer, and count the number of particles in a leukocyte area in the scatter diagram of the second leukocyte measurement system to obtain a second initial leukocyte count value;
the lipid particle number acquisition module comprises:
a lipid particle counting unit for generating a scatter diagram according to forward scattered light signals and side scattered light signals of a blood sample to be measured collected by a first white blood cell measurement system of a cell analyzer, identifying lipid particles based on the scatter diagram, and acquiring a first lipid particle count value or a second lipid particle count value; the first lipid particle count value is the number of lipid particles in the non-angiographic region; the second lipid particle count value is the number of lipid particles in the leukocyte region;
the correction module is used for subtracting the first lipid particle counting value from the first initial white blood cell counting value to obtain a corrected white blood cell counting value; or for subtracting the second lipid particle count value from the second initial leukocyte count value to obtain a corrected leukocyte count value.
7. The white blood cell counting device according to claim 6,
the scattered light signals collected by the second white blood cell measurement system comprise forward scattered light signals and side scattered light signals, and the particle number in the non-ghost area or the particle number in the white blood cell area is counted based on the two-dimensional scatter diagram of the forward scattered light signals and the side scattered light signals; and
the light signals collected by the first leukocyte measurement system include at least forward scattered light signals and side scattered light signals, and lipid particles are identified based on a two-dimensional scattergram of the forward scattered light signals and the side scattered light signals.
8. The white blood cell counting device of claim 7, wherein the light signals collected by the first white blood cell measurement system further include fluorescence signals, and wherein lipid particles are identified based on the three-dimensional scattergram of forward scattered light signals, side scattered light signals, and fluorescence signals.
9. The white blood cell counting device according to claim 6 or 7, further comprising:
a labeling module for labeling the lipid particles differently from the labeled leukocytes in the scatter plot of the second leukocyte measurement system.
10. The white blood cell counting apparatus of claim 6 or 7, further comprising an identification module for identifying lipid particles of a scatter plot of a second white blood cell measurement system, the identification module comprising:
the first area dividing unit is used for dividing a scatter diagram of the first white blood cell measuring system into a plurality of first sub-areas along the direction of lateral scattered light;
the second area dividing unit is used for dividing a scatter diagram of the second white blood cell measuring system into a plurality of second sub-areas along the direction of lateral scattered light;
the first region dividing unit and the second region dividing unit divide the sub-regions in the same way, so that each first sub-region and each second sub-region have one-to-one correspondence relationship;
the counting unit is used for counting the number of the lipid particles in the first subregion;
the identification unit is used for selecting the particles in the second sub-area and identifying the particles as lipid particles, so that the number of the lipid particles in the first sub-area is the same as that of the corresponding lipid particles in the second sub-area; alternatively, the ratio of the number of lipid particles in each first subregion to the number of lipid particles in the corresponding second subregion is the same.
11. A cell analyzer, characterized by comprising:
the conveying equipment is used for conveying the sample liquid to be detected to the optical detection equipment;
the optical detection device is used for irradiating the detected sample liquid flowing through the detection area, collecting various optical signals generated by the cells due to the irradiation of the light, and converting the optical signals into corresponding electric signals for output;
a white blood cell counting device according to any one of claims 6 to 10, for receiving and processing the electrical signal output by the optical detection means.
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| CN109490175A (en) * | 2018-10-17 | 2019-03-19 | 南宁优康电子科技有限公司 | Cell count sorter |
| WO2020133285A1 (en) * | 2018-12-28 | 2020-07-02 | 深圳迈瑞生物医疗电子股份有限公司 | Blood cell parameter correction method, blood sample detector and storage medium |
| WO2020252692A1 (en) * | 2019-06-19 | 2020-12-24 | 深圳迈瑞生物医疗电子股份有限公司 | Cell analyzer, impedance method-based white blood cells classification method, and computer-readable storage medium |
| CN117501127A (en) * | 2021-12-22 | 2024-02-02 | 深圳迈瑞动物医疗科技股份有限公司 | Sample analysis device and sample analysis method |
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