Improved method for dyeing immunofluorescence cell

A technology of immunofluorescence and staining method, which is applied in the preparation of test samples, instruments, measuring devices, etc., can solve the problems of time-consuming, cumbersome operation, and high cost, and achieve shortened time, reduced staining and washing steps, and easy operation Effect

Inactive Publication Date: 2010-10-20
ARMY MEDICAL UNIV
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Problems solved by technology

[0011] This method is a two-step multiple staining method, which first stains the cell surface antigen and then stains the antigen in the nucleus. It has a good staining effect and can accurately identify Treg cells, but there are cumbersome, time-consuming and costly procedures. shortcoming

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  • Improved method for dyeing immunofluorescence cell
  • Improved method for dyeing immunofluorescence cell
  • Improved method for dyeing immunofluorescence cell

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Embodiment Construction

[0026] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0027] In this example, the improved immunofluorescent cell staining method of the present invention was used to stain the spleen lymphocytes of Balb / c mice for CD4 and Foxp3, and then detected by flow cytometry, and compared with the method of eBioscience Company.

[0028] Experimental animals: 3 Balb / c mice aged 6-8 weeks, provided by the Experimental Animal Center of Third Military Medical University.

[0029] Experimental reagents: fluorescein isothiocyanate (FITC)-labeled rat anti-mouse CD4 antibody (FITC-CD4), allophycocyanin (APC)-labeled rat anti-mouse Foxp3 antibody (APC-Foxp3), isotype Control: FITC-labeled rat anti-mouse IgG2a (FITC-IgG2a), APC-labeled rat anti-mouse IgG2a (APC-IgG2a), and fixation / permeabilization solution, permeabilization solution, flow cytometry staining solution A product of eBioscience.

[0030] Experi...

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Abstract

The invention discloses an improved immune fluorescyte staining method, comprising 5 steps: fixing / permeating, permeating, surface staining, staining in nucleus, washing and weight dropping; the method of the invention is an improvement to the Foxp3 immune fluorescyte staining method of eBioscience Company, changes a two-step multiple staining method into a one-step multiple staining method, and carries out the staining on the surface of the cell and in the nucleus simultaneously, thus having the advantages of simple and convenient operation, quick staining, saving reagent, and reducing the cost, etc. The research shows that the method of the invention has no remarkable difference with the method of that of the eBioscience Company, can be used for the staining on the surface of the cell and in the nucleus simultaneously and has extremely good application prospect and promotional value.

Description

technical field [0001] The invention relates to the biological field, in particular to an improved immunofluorescent cell staining method. Background technique [0002] Regulatory T cells (Treg) are a type of T cells with immunosuppressive effects, which are an important part of the body's maintenance of self-tolerance, and play an important role in immune-related diseases, tumor immunity, and organ transplantation immunity. . At present, the expression level and clinical significance of peripheral blood Treg cells in patients with immune-related diseases, tumors and organ transplantation have become the focus of research. Studies have shown that the increase in the proportion of Treg cells may be one of the causes of immunosuppression in systemic lupus erythematosus, rheumatoid arthritis, lung cancer, gastric cancer, ovarian cancer, nasopharyngeal cancer, and hepatitis B virus infection; Treg cells in patients before and after treatment Quantitative changes can be used as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30G01N33/533
Inventor 倪兵赵韧吴玉章许桂莲王庆红付小岚贾正才王靖雪李健石晶磊黄泽民田易田志强
Owner ARMY MEDICAL UNIV
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