Application of flow cytometer in cell counting and method

A flow cytometer and cell counting technology, applied in scientific instruments, particle and sedimentation analysis, measuring devices, etc., can solve the problems of increasing the economic cost of detection, small share, and the inability to know the exact or absolute sample volume injected. , to overcome the heavy dependence on special counting instruments and consumables and personnel, meet clinical needs, and facilitate rapid detection

Pending Publication Date: 2022-04-12
北京海思特医学检验实验室有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional counting methods have different disadvantages. For example, the counting plate method is manual counting, which takes a long time and has high requirements for personnel. It takes a period of training before personnel can meet the requirements of cell counting, so the total economic and labor costs are high. The blood cell counting method takes a long time, and the instruments and consumables are expensive, which increases the economic cost of detection; while the flow cytometer with its own cell counting function has a small market share, and the flow cytometer with a large market share has a large market share due to its Volume delivery control is not absolute, i.e. the sample loading volume is consistent every time within the same instrument, but the exact or absolute sample volume injected cannot be known, thus resulting in these flow cytometers not capable of cell counting function, the density of the specimen or cell suspension to be tested cannot be known

Method used

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  • Application of flow cytometer in cell counting and method
  • Application of flow cytometer in cell counting and method
  • Application of flow cytometer in cell counting and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 Cell counting of the present invention compares with conventional counting plate counting method detection accuracy

[0068] The consistency of different acquisition speeds and acquisition times of the present invention and counting plate counts for the detection results of the same specimen were compared respectively.

[0069] Take 6 cases of clinical samples (clinical samples are bone marrow or peripheral blood of patients), including acute myeloid leukemia samples (sample number #1), acute lymphoblastic leukemia samples (sample number #2), chronic myeloid cell Leukemia specimen (sample number #3), myelodysplastic syndrome specimen (sample number #4), neuroblastoma specimen (sample number #5), myeloproliferative neoplasm specimen (sample number #6), Count according to the following methods:

[0070] 1) Sample preparation: Take 10 μl of the aforementioned specimen, put it into 390 μl of red blood cell lysate (commercial reagent, manufacturer: Lianke Bio, ...

Embodiment 2

[0095] Example 2 Application of Flow Cytometry to Count and Compare Between Different Personnel

[0096] Compare the consistency of counting results of the same specimen by different experimenters.

[0097] Take 10 cases of clinical samples, including acute myeloid leukemia (sample number #7-8), acute lymphoblastic leukemia (sample number #9-10), chronic myeloid leukemia (sample number #11-12 ), myelodysplastic syndrome (sample number #13-14), neuroblastoma (sample number #15), myeloproliferative neoplasms (sample number #16). For the cell counting method, refer to the operation method in Example 1. The test results are as follows:

[0098] Table 2: Comparison results among different personnel for counting by flow cytometry

[0099] Specimen number Person A Person B CV value (100%) #7 31.6 29.8 4.1 #8 26.1 26.9 2.1 #9 187.5 193.4 2.2 #10 46.5 48.5 3.0 #11 158.1 166.8 3.8 #12 252.2 274.6 6.0 #13 119.3 125 3.3 ...

Embodiment 3

[0101] The influence of embodiment 3 different erythrocyte lysate volumes on counting result

[0102] The present invention uses a specimen volume of 10 μl, and compares the volumes of different erythrocyte lysates. If the volume of the cell suspension is too small, the volume of the suspension cannot meet the requirement of obtaining the cells for 30 seconds after the liquid flow stabilizes, so start the experiment with 190 μl of lysate. However, if the volume of the cell suspension is too large, the cell concentration will be small, the number of cells obtained will be small, and the boundary between cells and debris will not be clear. Therefore, the effects of 190μl, 290μl, 390μl and 490μl of erythrocyte lysate on the counting results were compared respectively. Other steps are with embodiment 1.

[0103] 6 clinical specimens were taken, including acute myeloid leukemia specimens (sample number #1), acute lymphoblastic leukemia specimens (sample number #2), chronic myeloi...

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Abstract

The invention relates to the technical field of biology, in particular to application and a method of a flow cytometer in cell counting, a liquid flow system of the flow cytometer is a liquid flow system for sampling by using pressure difference between a sample and sheath fluid, and the cell counting comprises the step of detecting the total cell number or cell concentration in a sample to be detected, the specimen to be detected is selected from peripheral blood or bone marrow or a suspension of a cell line. According to the cell counting method, the detection speed can be greatly increased, the detection accuracy is high, the sensitivity is high, the use amount of samples is small, a microscope and a cell counting instrument do not need to be additionally purchased, and cell counting and flow online detection can be completed by using one flow cytometer; meanwhile, long-time training on experimenters due to manual counting is omitted, serious dependence on special counting instruments and consumables and personnel is overcome, rapid detection of clinical samples is facilitated, patients can be diagnosed and treated in time, and clinical requirements are met.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use and method of flow cytometry in cell counting. Background technique [0002] At present, the methods of cell counting mainly include the counting plate method, the blood cell counter method, and a small number of flow cytometers with built-in cell counting functions. Conventional counting methods have different disadvantages. For example, the counting plate method is manual counting, which takes a long time and has high requirements for personnel. It takes a period of training before personnel can meet the requirements of cell counting, so the total economic and labor costs are high. The blood cell counting method takes a long time, and the instruments and consumables are expensive, which increases the economic cost of detection; while the flow cytometer with its own cell counting function has a small market share, and the flow cytometer with a large market share has a large ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14
Inventor 刘丽媛关天阳周雪邵萌王显凤吴纯斌倪花丽张娟程宇梁超陈忠黄士昂
Owner 北京海思特医学检验实验室有限公司
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