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Method for assaying T-lymphocyte subpopulations of swine peripheral blood by flow cytometry

A technology of flow cytometry and lymphocytes, which is applied in the field of flow cytometry to detect porcine peripheral blood T lymphocyte subsets, can solve the problems of large fluctuations and instability of test results, achieve less cell fragments, avoid errors, and improve Effects of Authenticity and Objectivity

Inactive Publication Date: 2017-09-15
成都扬克斯生物科技有限公司
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Problems solved by technology

[0003] In view of this, the present invention provides a flow cytometry method for the detection of porcine peripheral A method for detecting T lymphocyte subsets in blood, establishing a stable and standardized detection method for T lymphocyte subsets in porcine peripheral blood

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  • Method for assaying T-lymphocyte subpopulations of swine peripheral blood by flow cytometry
  • Method for assaying T-lymphocyte subpopulations of swine peripheral blood by flow cytometry
  • Method for assaying T-lymphocyte subpopulations of swine peripheral blood by flow cytometry

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Embodiment

[0057] To detect the expression of CD3 / CD4 / CD8 in the fresh peripheral blood of piglets infected with a certain respiratory virus by intranasal drip.

[0058] The 60-day-old test pigs were equally divided into two groups. The pigs in the control group were sprayed with saline nasal cavity, and the pigs in the challenge group were sprayed with virus liquid in the nasal cavity. After feeding for 21 days, 6 pigs were randomly selected in each group, and blood was collected from the jugular vein to prepare anticoagulant blood, which was used to detect the distribution of T lymphocyte subsets in the peripheral blood of the pigs.

[0059] Step 1, take 2mL of peripheral venous blood from the tested pig, put it in a 2mL EDTA-Na2 anticoagulant tube, vortex and mix immediately, and prepare anticoagulant blood;

[0060] Step 2: Pipette 100 μL of the anticoagulated blood into two flow tubes, one tube was stored at 4°C for later use; the other tube was added with mouse anti-pig CD3-SPRD (S...

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Abstract

The invention discloses a method for assaying T-lymphocyte subpopulation of swine peripheral blood by flow cytometry. The method is characterized by including the following steps: swine peripheral venous blood is extracted to prepare anticoagulation blood; the anticoagulation blood is sucked, anti-swine CD3, CD4 and CD8 monoclonal antibodies are respectively added, vortex mixing is carried out, and dyeing is carried out in the dark at 4 DEG C; a red blood cell lysis buffer is used for rupturing red blood cells; a PBS solution is used for washing and resuspending cells, and a flow cytometer is used for assaying; and an assay result is analyzed, so that a proportion of the T-lymphocyte subpopulations of the swine peripheral blood is obtained. the stable and standard method for assaying the T-lymphocyte subpopulations of the swine peripheral blood by flow cytometry is successfully established; by improving for the problems of target cell choice, compensative regulation and parameter selection in the process of pretreating, assaying and analyzing a sample, errors caused by human factors in the process of an experiment are prevented, and thereby the authenticity and objectivity of result judgment are increased.

Description

technical field [0001] The invention belongs to the technical field of cell detection, in particular to a method for detecting porcine peripheral blood T lymphocyte subsets by flow cytometry. Background technique [0002] Flow cytometry (Flow Cytometry, FCM) is a new technique for fluorescent quantitative analysis of chemical components on the surface of a single cell or in a cell. Its working principle is to perform multi-parameter quantitative analysis after fluorescently labeling a single cell or molecule. As a rapid cell analysis technology, this technology is widely used in basic medical research fields such as immunology and cell biology. Flow cytometry can realize the detection of T lymphocyte subsets by identifying specific immunofluorescence signals on the cell surface, which is helpful for evaluating the cellular immune function of the body. In veterinary clinical and scientific research, it is found that sample pretreatment, parameter adjustment during detection...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14
CPCG01N15/1404G01N15/1434
Inventor 彭西于正强胡东李林蔚
Owner 成都扬克斯生物科技有限公司
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