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A method for detecting phagocyte function based on flow cytometry

A technology of flow cytometry and phagocytic cells is applied in the field of evaluating the ability of phagocytic cells to phagocytose bacteria based on flow cytometry, which can solve the problems of unfavorable promotion and use, difficult to distinguish detection, weak FITC fluorescence, etc. Simple and easy cost, good repeatability

Active Publication Date: 2021-12-21
SHANDONG UNIV
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Problems solved by technology

At present, there are not many reports on the detection of phagocytic function of phagocytic cells. It has been reported to use fluorescein FITC-labeled bacteria prepared in carbonate buffer to detect phagocytic function of phagocytic cells. However, this method has the disadvantages of weak fluorescence of FITC and difficult to distinguish detection. Not conducive to promotion

Method used

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  • A method for detecting phagocyte function based on flow cytometry
  • A method for detecting phagocyte function based on flow cytometry

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Embodiment 1

[0037] 1. Take BALB / c mouse peritoneal macrophages, count them, take 1 million cells, inoculate them in a cell culture plate and cultivate them; take BALB / c mouse bone marrow cells, and use Percoll density gradient centrifugation to separate neutrophils. Counting takes a certain number of cells.

[0038] 2. Add 100 μL of thiazole orange staining solution to the E. coli bacterial liquid, the concentration of the thiazole orange staining solution is 0.5 mg / mL, shake and incubate at 37°C for 60 minutes, wash, centrifuge, and discard the supernatant;

[0039] 3. Add 5ml of PBS to the bacterial pellet at the bottom after centrifugation, and count and analyze by flow cytometry;

[0040] 4. Add 200 μL of Escherichia coli liquid stained with thiazole orange to the phagocyte culture medium, and incubate for 20 minutes;

[0041] 5. Digest and collect phagocytes, add 0.1ml PBS, and analyze by flow cytometry, the excitation light of thiazole orange is 488nm, and the emission light is 530...

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Abstract

The invention discloses a method for detecting phagocyte function based on flow cytometry. In the method of the invention, Escherichia coli bacteria liquid and thiazole orange staining solution are shaken and incubated on a constant temperature shaker for an appropriate time, washed with PBS to remove non-specific adhesion, and centrifuged. Discard the supernatant. Finally, add an appropriate amount of PBS to resuspend, add to the phagocyte culture medium, and incubate in the incubator for an appropriate time. Phagocytes were collected by digestion, and the cell distribution of fluorescent populations was quantitatively analyzed on a flow cytometer, thereby analyzing the function of phagocytes to capture phagocytic bacteria. The invention is fast, accurate, simple and easy to implement, and has low cost, which is beneficial to popularization and application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for evaluating the phagocytosis ability of phagocytes based on flow cytometry. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Phagocytes are a group of cells with phagocytic function in the body, mainly including mononuclear phagocyte system and neutrophils. The mononuclear phagocyte system includes monocytes free in the blood and macrophages developed after entering various tissues. Among them, macrophages have a strong phagocytic ability and are also a major class of antigen-presenting cells, which play a key role in the induction and regulation of specifi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14G01N21/64
CPCG01N15/1434G01N21/6402
Inventor 赵云雪荆卫强王甘雨毕玉璇韩丽辉安杰郭兴
Owner SHANDONG UNIV
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