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Method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry

A technology of flow cytometry and lymphocytes, which is applied in the field of flow cytometry detection of chicken peripheral blood T lymphocyte subsets, can solve the problems of large fluctuations and instability of test results, improve authenticity and objectivity, and avoid effect of error

Inactive Publication Date: 2017-08-18
CHINA WEST NORMAL UNIVERSITY
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Problems solved by technology

[0004] In view of this, the present invention provides a flow cytometry method for detection of chicken peripheral A method for detecting T lymphocyte subsets in blood, establishing a stable and standardized detection method for T lymphocyte subsets in chicken peripheral blood

Method used

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  • Method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry
  • Method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry
  • Method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry

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Embodiment

[0060] Detection of oral exposure to AFB 1 The expression of CD3 / CD4 / CD8 in fresh peripheral blood of chicks.

[0061] The test chickens were equally divided into two groups, and the chicks in the control group ate the normal control diet, AFB 1 Chicks in the group fed with a certain dose level of AFB 1 daily ration. After 21 days of feeding, 6 chickens 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 chickens.

[0062] Step 1, first add 1mL of lymphocyte separation medium to the flow tube, and then carefully add an equal volume of fresh blood, so that the fresh blood is completely located on the upper layer of the lymphocyte separation medium to avoid mixing the two;

[0063] Step 2, place the flow tube in an ordinary high-speed centrifuge, and centrifuge at a speed of 2500r / min for 15min;

[0064] Step 3. Afte...

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Abstract

The invention discloses a method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry; the method comprises the steps of collecting avian peripheral venous blood, preparing anticoagulant blood, and treating with lymphocyte separate to obtain peripheral blood leukocytes; centrifugally washing the peripheral blood leukocytes with PBS (phosphate buffer solution) to prepare single-cell suspension; sucking the single-cell suspension, adding anti-chicken CD3, CD4 and CD8 monoclonal antibodies, mixing well by cyclone, and staining in the shade at 4 DEG C; after staining, washing with PBS, resuspending cells, and detecting with a flow cytometer; analyzing detection results, the ratio of avian peripheral blood T lymphocyte subsets. The method according to the invention which detects avian peripheral blood T lymphocyte subsets stably and regularly is established, thereby solving the problem that differences between sample pretreatment, adjustment of voltage and fluorescence compensation upon detection and processing modes for result analysis cause high fluctuations of detection results and instability.

Description

technical field [0001] The invention belongs to the technical field of cell detection, and in particular relates to a method for detecting chicken peripheral blood T lymphocyte subsets by flow cytometry. Background technique [0002] Flow cytometry (Flow Cytometry, FCM) is a new technology for quantitative analysis and sorting of chemical components on the surface or intracellular of a single cell. FCM mainly separates cells with different cell sizes and intracellular granularity by collecting forward scattered light (FSC) and side scattered light (SSC). And by identifying specific fluorescent markers on the cell surface and inside the cells, the detection of DNA, RNA, cytokines, cell surface antigens, etc. is realized. As a rapid cell analysis technology, this technology is not only used in basic medical research such as immunology and microbiology, but also widely used in clinical testing such as oncology and hematology. [0003] Flow cytometry can realize the detection ...

Claims

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Application Information

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IPC IPC(8): G01N15/14
CPCG01N15/1425
Inventor 彭西张泽钧袁施彬吴邦元
Owner CHINA WEST NORMAL UNIVERSITY
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