CN115406816A - Method for measuring reticulocyte, sample analyzer and storage medium - Google Patents
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- 210000001995 reticulocyte Anatomy 0.000 title claims abstract description 137
- 238000000034 method Methods 0.000 title claims abstract description 21
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- 238000001514 detection method Methods 0.000 claims abstract description 109
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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1486—Counting the particles
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Abstract
The present application provides a measurement method of reticulocytes, including: acquiring a first sample and a second sample, wherein the first sample and the second sample are homologous samples; carrying out first pretreatment on the first sample through a reticulum reagent to obtain a first sample to be detected; enabling a first sample to be detected to pass through an optical detection channel of the first flow cytometry detection assembly by using the first flow cytometry detection assembly so as to obtain the number of reticulocytes; passing the second sample through a detection channel of the cell detection assembly to obtain the number of red blood cells by using the cell detection assembly; parameters of reticulocytes were obtained based on the number of reticulocytes and the number of erythrocytes. In the measuring method, the reticulocytes are counted through the optical detection channel, and the erythrocytes are counted through the detection channel of the cell detection assembly, so that the accuracy of parameter measurement of the reticulocytes can be improved.
Description
Technical Field
The present application relates to the field of medical devices, and in particular, to a method for measuring reticulocytes, a sample analyzer, and a storage medium.
Background
Reticulocytes are not fully mature red blood cells, and their value in the peripheral blood reflects the production function of bone marrow erythrocytes, thus being of great significance for diagnosis of hematological disorders and observation of therapeutic responses.
At present, products for testing reticulocytes basically stain the reticulocytes after spheroidization of the erythrocytes and the reticulocytes; a graph with red blood cells and reticulocytes is obtained, and the algorithm calculates the percentage of reticulocytes by obtaining counts of red blood cells and reticulocytes after segmentation according to position.
However, the measurement method of reticulocytes may be relatively dependent on the processing capability of the algorithm, and the counts of the obtained erythrocytes and reticulocytes may have a deviation, resulting in inaccurate test results.
Disclosure of Invention
The application provides a measurement method of reticulocytes, a sample analyzer and a storage medium, which aim to solve the technical problem that in the prior art, the measurement method of reticulocytes is relatively dependent on the processing capacity of an algorithm, so that the accuracy of a measurement result is not high.
In order to solve the technical problem, the application adopts a technical scheme that: there is provided a measurement method of reticulocytes, the measurement method including: acquiring a first sample and a second sample, wherein the first sample and the second sample are homologous samples; carrying out first pretreatment on the first sample through a reticulum reagent to obtain a first sample to be detected; enabling a first sample to be detected to pass through an optical detection channel of the first flow cytometry detection assembly by using the first flow cytometry detection assembly so as to obtain the number of reticulocytes; passing the second sample through a detection channel of the cell detection assembly to obtain the number of red blood cells by using the cell detection assembly; parameters of reticulocytes were obtained based on the number of reticulocytes and the number of erythrocytes.
Further, performing first pretreatment on the first sample by using a reticuloendothelial reagent to obtain a first sample to be detected, wherein the first pretreatment comprises the steps of adding the reticuloendothelial reagent and the first sample into a first sample preparation assembly and mixing to obtain a sample mixed solution; and incubating the sample mixed solution at a preset first temperature for a preset time to obtain a first sample to be detected.
Further, incubating the sample mixture at a predetermined first temperature for a predetermined time to obtain a first sample to be tested, including incubating the sample mixture at a temperature of 25-45 ℃ for 3-10 minutes to ghost or lyse red blood cells in the sample mixture by the reticulocyte reagent, and staining nucleic acids of the reticulocytes to obtain the first sample to be tested.
Further, adding the reticule red reagent and the first sample into the first sample preparation assembly for mixing to obtain a sample mixed solution, wherein the reticule red reagent is heated to a preset second temperature; and mixing the heated reticulum reagent and the first sample in a first sample preparation assembly to obtain a sample mixed solution.
Further, the first sampling module comprises: one of a WBC pool, RBC pool, HGB pool and mixing pool.
Further, the reticulocytes include high nucleic acid reticulocytes and low nucleic acid reticulocytes, and the parameters of the reticulocytes include: one or more of reticulocytes per cent of the number of red blood cells, low nucleic acid reticulocytes per cent of the number of red blood cells, high nucleic acid reticulocytes per cent of the number of red blood cells, reticulocyte number, low nucleic acid reticulocyte number, and high nucleic acid reticulocyte number.
Further, the measuring method further comprises: carrying out second pretreatment on the second sample to obtain a second sample to be detected, and enabling the second sample to pass through a detection channel of the cell detection assembly by using the cell detection assembly to obtain the number of the red blood cells, wherein the second pretreatment comprises the following steps: and (3) enabling the second sample to be detected to pass through a detection channel of the cell detection assembly by using the cell detection assembly so as to obtain the number of the red blood cells.
Further, using the cell detection assembly, passing the second sample through the detection channel of the cell detection assembly to obtain the number of red blood cells, comprising: the second sample is passed through the impedance detection channel of the impedance detection assembly to obtain the number of red blood cells using the impedance detection assembly, or the second sample is passed through the optical detection channel of the second flow cytometry assembly to obtain the number of red blood cells using the second flow cytometry assembly.
In order to solve the technical problem, the other technical scheme adopted by the application is as follows: there is provided a sample analyzer comprising: the flow cell detection device comprises a first flow cell detection assembly, a cell detection assembly and a controller, wherein the controller is connected with the first flow cell detection assembly and the cell detection assembly, and the controller is used for realizing the measurement method of the reticulocytes of any embodiment.
In order to solve the technical problem, the other technical scheme adopted by the application is as follows: there is provided a computer readable storage medium having stored thereon program instructions which, when executed by a processor, implement the method of measuring reticulocytes as in any one of the above embodiments.
The beneficial effect of this application is: in contrast to the prior art, the measurement method of reticulocytes of the present application includes: obtaining a first sample and a second sample; carrying out first pretreatment on the first sample through a reticulocyte reagent to obtain a first sample to be detected; enabling a first sample to be detected to pass through an optical detection channel of the first flow cytometry detection assembly by using the first flow cytometry detection assembly so as to obtain the number of reticulocytes; passing the second sample through a detection channel of the cell detection assembly to obtain the number of red blood cells by using the cell detection assembly; parameters of reticulocytes were obtained based on the number of reticulocytes and the number of erythrocytes. In this application, handle the sample through the reticulocyte reagent, get rid of the interference of erythrocyte at optical channel to the reticulocyte count to distinguish the reticulocyte, come to count the reticulocyte through optical channel, the rethread detection channel carries out the erythrocyte count, so, can avoid the error that the algorithm processing brought, improve the accuracy of reticulocyte and erythrocyte detection count, thereby improve the accuracy that the relevant parameter of reticulocyte measures.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without inventive efforts, wherein:
FIG. 1 is a schematic flow chart diagram illustrating an embodiment of a method for measuring reticulocytes provided herein;
FIG. 2 is a schematic block diagram of an embodiment of a measurement method for reticulocytes provided by the present application;
FIG. 3 is a schematic flow chart of one embodiment of step S12 in FIG. 1;
FIG. 4 is a block diagram of one embodiment of a sample analyzer provided herein;
FIG. 5 is a block diagram of an embodiment of a computer-readable storage medium provided herein.
Detailed Description
The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without making any creative effort belong to the protection scope of the present application.
It should be noted that if directional indications (such as upper, lower, left, right, front, rear, 8230; \8230;) are referred to in the embodiments of the present application, the directional indications are only used for explaining the relative positional relationship between the components in a specific posture (as shown in the attached drawings), the motion situation, etc., and if the specific posture is changed, the directional indications are correspondingly changed.
In addition, if there is a description of "first", "second", etc. in the embodiments of the present application, the description of "first", "second", etc. is for descriptive purposes only and is not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the feature. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present application.
The application provides a method for measuring reticulocytes, which can accurately count reticulocytes and red blood cells in a sample, thereby improving the accuracy of reticulocyte related parameters.
Referring to fig. 1 and fig. 2, fig. 1 is a schematic flow chart of an embodiment of a measurement method of reticulocytes provided in the present application, and fig. 2 is a schematic frame diagram of an embodiment of the measurement method of reticulocytes provided in the present application, the measurement method including:
s11: a first sample and a second sample are obtained, the first sample and the second sample being homologous samples.
When measuring reticulocytes of a sample, a first sample and a second sample are obtained. In particular, the sample analyzer may acquire the first sample and the second sample through the sampling assembly. Wherein the first sample and the second sample are homologous samples. The first and second samples are homologous blood samples to be measured. The first sample is used for counting and detecting reticulocytes, and the second sample is used for counting and detecting erythrocytes.
For example, the sample analyzer includes a sampling assembly, and the sample analyzer can draw multiple blood samples through the sampling assembly and add the multiple blood samples to the first sample preparation assembly and the second sample preparation assembly respectively to obtain a first sample for reticulocyte counting and a second sample for red blood cell counting.
The first sample preparation assembly and the second sample preparation assembly may include one of a WBC (white blood cell) pool, a RBC (red blood cell) pool, an HGB (hemoglobin) pool, and a mixing pool, as long as the first sample preparation assembly and the second sample preparation assembly can be used for uniformly mixing and preparing samples. The first sample preparation assembly and the second sample preparation assembly can be the same assembly or different assemblies.
S12: and carrying out first pretreatment on the first sample through the reticulum reagent to obtain a first sample to be tested.
And performing first pretreatment on the first sample through a reticulocyte reagent, wherein the reticulocyte reagent can be used for dyeing the nucleic acid of the reticulocyte in the first sample, and removing the interference of the erythrocyte on reticulocyte counting in an optical channel so as to distinguish the reticulocyte, so that the obtained first sample to be detected can be used for counting the reticulocyte.
Furthermore, a specific reticulocyte reagent can be selected, which is not only used for hemographing or lysing the red blood cells in the first sample, but also used for staining the nucleic acid of the reticulocyte and removing the red blood cells in the first sample as much as possible to obtain the first sample to be tested. By the method, the influence of the erythrocytes on measurement of the reticulocytes can be reduced, and the accuracy of the detection result is improved.
In one embodiment, as shown in FIG. 3, the step of subjecting the first sample to a first pretreatment with a reticulocyte reagent to obtain a first sample to be tested comprises:
s121: and adding the reticulum reagent and the first sample into the first sample preparation assembly for mixing to obtain a sample mixed solution.
Specifically, a reticular reagent and a first sample may be added to the first sample preparation assembly and mixed to obtain a sample mixture. Further, the reticulocyte reagent can be preheated to raise the temperature of the reticulocyte reagent to a preset second temperature, then the heated reticulocyte reagent is added into the first sample preparation assembly, then the first sample is added, and the first sample and the heated reticulocyte reagent are mixed to obtain a sample mixed solution. For example, the reticulant reagent may be heated to 30 degrees before being added to the first sample preparation assembly and mixed with the first sample. In this way, the effect of the reticular reagent on the first sample can be improved.
S122: and incubating the sample mixed solution at a preset first temperature for a preset time to obtain a first sample to be detected.
After obtaining the mixed solution of the reticulocyte reagent and the first sample, incubating the mixed solution of the samples at a preset first temperature for a preset time so that the reticulocyte reagent can remove red blood cells as much as possible. Through incubating the sample mixed liquid, the influence of the red blood cells on the reticulocyte counting can be reduced, and the accuracy of the reticulocyte counting can be improved.
Further, the sample mixture may be incubated at a temperature ranging from 25 to 45 ℃ for 3 to 10 minutes to hemolyze or lyse the red blood cells in the sample mixture by the reticulocyte reagent and stain the nucleic acids of the reticulocytes to obtain the first sample to be measured. For example, the sample mixture may be incubated at 30 degrees for 5 minutes to form a first test sample.
Optionally, in some embodiments, a second pretreatment may also be performed on the second sample to obtain a second test sample, so that the second test sample can be used for counting red blood cells. For example, the second sample may be diluted to obtain the second sample to be measured, or the second sample may be heated to obtain the second sample to be measured.
In a specific embodiment, the diluent and the second sample may be added to the second sample preparation assembly, for example, the diluent and the second sample are added to the RBC pool to be mixed and diluted, so as to obtain a second sample to be tested.
Alternatively, the second sample may be directly detected without preprocessing.
S13: and (3) enabling the first sample to be detected to pass through the optical detection channel of the first flow cytometry detection assembly by using the first flow cytometry detection assembly so as to obtain the number of the reticulocytes.
After the first sample to be detected is obtained, the first sample to be detected passes through the optical detection channel of the first flow cytometry detection assembly by utilizing the first flow cytometry analysis technology so as to count the reticulocytes. Reticulocytes include high nucleic acid reticulocytes and low nucleic acid reticulocytes. The number of reticulocytes, the number of high nucleic acid reticulocytes, and the number of low nucleic acid reticulocytes can be obtained by the first flow cytometry assembly.
The flow cytometry analysis technology is a single cell quantitative analysis and sorting technology which is carried out by utilizing a flow cytometer. The first flow cell detection assembly mainly comprises a light source, a flow channel (i.e. an optical detection channel) and a signal detection system.
After the first sample to be detected is obtained, the first sample to be detected is optically detected through a flow cytometry analysis technology, and specifically, the first sample to be detected can pass through an optical detection channel through the first conveying assembly so as to count the reticulocytes. For example, the first sample to be measured may be passed through the DIFF channel, and then the first sample to be measured passing through the DIFF channel may be detected by the light source and the signal detection system to count reticulocytes.
The counting of particles by flow cytometry techniques and flow cytometry components is a category understood by those skilled in the art and will not be described in detail herein.
S14: and (3) utilizing the cell detection assembly, enabling the second sample to pass through a detection channel of the cell detection assembly so as to obtain the number of the red blood cells.
After the second sample is obtained, the second sample can pass through a detection channel of the cell detection assembly through the cell detection assembly to obtain the number of the red blood cells. Specifically, the second sample may be subjected to a measurement of the number of red blood cells by impedance detection or flow cytometry.
Optionally, the second sample may be preprocessed to obtain the second sample to be detected, for example, the second sample is diluted, and the like, specifically refer to the description in the step S12, and then the second sample to be detected passes through the detection channel of the cell detection assembly to count the red blood cells.
Alternatively, the red blood cells in the second test sample can be counted by an impedance detection method using the impedance detection component. And in the process of impedance detection, enabling the second sample to be detected to pass through the impedance detection channel. For example, the impedance detection assembly includes an impedance detection cell, which may be a WBC cell or an RBC detection cell, where the WBC detection cell and the RBC detection cell each include a front cell and a rear cell, each front cell is open at an upper portion, a front cell electrode is installed in the front cell, a rear cell electrode is installed in the rear cell, and one end of each of the front cell electrode and the rear cell electrode is exposed on a surface of the cell body to form an electrode contact point. Microporous sheets for detecting the number of cells are arranged between the front pool and the rear pool; when the measurement is carried out, under the action of negative pressure, a second sample to be measured in the front pool enters the rear pool through the microporous sheet, and the second sample to be measured sequentially passes through the jewel holes of the microporous sheet so as to count red blood cells in the second sample to be measured.
Optionally, the second flow cytometry assembly can be used to count red blood cells in the second test sample by flow cytometry. In the detection process, a second sample to be detected passes through the optical detection channel of the second flow cytometry detection assembly to obtain the number of the red blood cells. In this way, the number of red blood cells can be accurately obtained.
Further, the second flow cell detection module and the first flow cell detection module may be the same module or different modules. When the second flow cell detection assembly and the first flow cell detection assembly are different assemblies, the quantity detection of the red blood cells and the quantity detection of the reticulocytes can be carried out simultaneously, so that the detection time is saved. When the second flow cell detection assembly and the first flow cell detection assembly are the same assembly, the quantity detection of the red blood cells and the quantity detection of the reticulocytes can be carried out according to time sequence, so that the structure of the instrument can be simplified, and the hardware cost is saved.
S15: parameters of reticulocytes were obtained based on the number of reticulocytes and the number of erythrocytes.
After obtaining the number of reticulocytes and the number of red blood cells, the parameters of reticulocytes can be obtained based on the above-described number of reticulocytes and the number of red blood cells. Wherein the reticulocytes comprise high nucleic acid reticulocytes and low nucleic acid reticulocytes, and the parameters of the reticulocytes comprise: one or more of reticulocytes, low nucleic acid reticulocytes, high nucleic acid reticulocytes, and high nucleic acid reticulocytes. For example, the percentage of reticulocytes in red blood cells can be obtained based on the number of reticulocytes and the number of red blood cells.
Among the measurement method of reticulocyte that this application provided, handle the sample through the reticulocyte reagent, get rid of the interference of erythrocyte at optical channel to the reticulocyte count, distinguish the reticulocyte, count the reticulocyte through optical channel, the rethread detection channel carries out the erythrocyte count, so, can avoid the error that the algorithm processing brought, improve the accuracy that reticulocyte and erythrocyte detected the count, thereby improve the accuracy that the relevant parameter of reticulocyte measures.
Fig. 4 is a schematic diagram of a sample analyzer according to an embodiment of the present disclosure. The sample analyzer 40 includes a first flow cell detection assembly 41, a cell detection assembly 42, and a controller 43, and the controller 34 is connected to the first flow cell detection assembly 41 and the cell detection assembly 42.
Optionally, the sample analyzer 40 may further include a sampling assembly (not shown), a first sample preparation assembly (not shown), and a second sample preparation assembly (not shown). The sample analyzer 40 may collect the first sample and the second sample via the sampling assembly. The first sample preparation assembly is used for receiving a first sample and preparing the first sample to obtain a first sample to be detected. The second sample preparation assembly is used for receiving a second sample and is used for preparing the second sample to obtain a second sample to be detected.
The controller 43 is configured to implement the steps of the measurement method of reticulocytes of any one of the above embodiments. Please refer to the description of any of the above embodiments for the steps of the measurement method of reticulocytes, which are not repeated herein.
The sample analyzer 40 can measure the parameters of the reticulocytes, and the accuracy of the measurement result is high.
Fig. 5 is a schematic diagram illustrating a frame of an embodiment of a computer-readable storage medium provided in the present application. The computer readable storage medium 50 stores program instructions 51 executable by the processor, the program instructions 51 being for implementing the steps of the method for measuring reticulocytes of any of the above embodiments.
The program instructions 51 may be stored in the computer-readable storage medium 50 in the form of a software product, and include several instructions to make a device or a processor execute all or part of the steps of the methods according to the embodiments of the present application.
The processor may also be referred to as a CPU (Central Processing Unit). The processor may be an integrated circuit chip having signal processing capabilities. The Processor may also be a general purpose Processor, a Digital Signal Processor (DSP), an Application Specific Integrated Circuit (ASIC), a Field Programmable Gate Array (FPGA) or other Programmable logic device, discrete Gate or transistor logic, discrete hardware components. A general purpose processor may be a microprocessor or the processor may be any conventional processor or the like. In addition, the processors may be collectively implemented by an integrated circuit chip.
The computer-readable storage medium 50 is a medium in a computer memory for storing some discrete physical quantity. Among them, the computer-readable storage medium 50 includes: a U disk, a removable hard disk, a Read-Only Memory (ROM), a Random Access Memory (RAM), a magnetic disk or an optical disk, and other various media capable of storing the program instruction 51 code.
The above embodiments are merely examples and are not intended to limit the scope of the present disclosure, and all modifications, equivalents, and flow charts using the contents of the specification and drawings of the present disclosure, which are directly or indirectly applied to other related technical fields, are included in the scope of the present disclosure.
Claims (10)
1. A method for measuring reticulocytes, comprising:
obtaining a first sample and a second sample, wherein the first sample and the second sample are homologous samples;
carrying out first pretreatment on the first sample through a reticulocyte reagent to obtain a first sample to be detected;
making the first sample to be detected pass through an optical detection channel of the first flow cell detection assembly by using the first flow cell detection assembly to obtain the quantity of the reticulocytes;
passing the second sample through a detection channel of a cell detection assembly to obtain the number of red blood cells;
obtaining a parameter of the reticulocytes based on the number of the reticulocytes and the number of the red blood cells.
2. The measurement method according to claim 1, wherein said first sample is subjected to a first pretreatment by a reticulocyte reagent to obtain a first sample to be measured, comprising,
adding the reticule red reagent and the first sample into a first sample preparation assembly for mixing to obtain a sample mixed solution;
and incubating the sample mixed solution at a preset first temperature for a preset time to obtain the first sample to be detected.
3. The method of claim 2, wherein incubating the sample mixture at a predetermined first temperature for a predetermined time to obtain the first sample to be tested comprises,
and incubating the sample mixed liquor for 3-10 minutes at the temperature range of 25-45 ℃ so as to enable the red blood cells in the sample mixed liquor to be ghosted or lysed through the reticulocyte reagent, and staining nucleic acids of the reticulocyte to obtain the first sample to be detected.
4. The method of claim 2, wherein the adding the reticule red reagent and the first sample to a first sample preparation assembly and mixing to obtain a sample mixture comprises,
heating the reticulocyte reagent to a preset second temperature;
mixing the heated reticulum reagent and the first sample in the first sample preparation assembly to obtain the sample mixed solution.
5. The measurement method of claim 2, wherein the first sample preparation assembly comprises: one of a WBC pool, a RBC pool, a HGB pool, and a mixing pool.
6. The method of measuring according to claim 1, wherein the reticulocytes comprise high nucleic acid reticulocytes and low nucleic acid reticulocytes,
the parameters of the reticulocytes include: one or more of the percentage of the number of red blood cells by reticulocytes, the percentage of the number of red blood cells by low nucleic acid reticulocytes, the percentage of the number of red blood cells by high nucleic acid reticulocytes, the number of low nucleic acid reticulocytes, and the number of high nucleic acid reticulocytes.
7. The measurement method according to claim 1, characterized in that the measurement method further comprises: performing second pretreatment on the second sample to obtain a second sample to be detected,
the using the cell detection assembly, passing the second sample through the detection channel of the cell detection assembly to obtain the number of red blood cells, including: and the cell detection assembly is utilized to enable the second sample to be detected to pass through a detection channel of the cell detection assembly so as to obtain the number of the red blood cells.
8. The method of measuring according to claim 1, wherein said passing the second sample through a detection channel of a cell detection assembly using the cell detection assembly to obtain the number of red blood cells comprises:
using an impedance detection component, making the second sample pass through an impedance detection channel of the impedance detection component to obtain the number of the red blood cells; or the like, or, alternatively,
and using a second flow cell detection assembly to make the second sample pass through an optical detection channel of the second flow cell detection assembly to obtain the number of the red blood cells.
9. A sample analyzer, comprising: a first flow cell assay assembly, a cell assay assembly, and a controller connecting the first flow cell assay assembly and the cell assay assembly, the controller being configured to implement the method of measuring reticulocytes of any one of claims 1-8.
10. A computer-readable storage medium having stored thereon program instructions, which when executed by a processor, implement the method of measuring reticulocytes of any one of claims 1 through 8.
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