Using 9-color Flow Cytometry Immunophenotyping to Detect Activation and Apoptosis of Human TCR Vβ Lymphocyte Subpopulations in Peripheral Blood Samples

To establish an assay using 9-color flow cytometry immunophenotyping to detect activation and apoptosis of human TCR Vβ lymphocyte subpopulations in peripheral blood samples. We used 5 antibodies (CD3, CD4, CD8, CD95, CD69), phospholipids binding proteins Annexin V, TCR Vβ Repertoire Kit and nucleus...

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Published inSichuan da xue xue bao. Journal of Sichuan University. Yi xue ban Vol. 47; no. 2; p. 262
Main Authors Huang, Qiao-rong, Wen, Bo, Li, Xue, Hu, Lian, Wan, Xue-mei, Zeng, Sheng-ping, Meng, Wen-tong
Format Journal Article
LanguageChinese
Published China 01.03.2016
Subjects
Antibodies
Apoptosis
Color
Flow Cytometry
Humans
Immunophenotyping - methods
Lymphocyte Activation
Lymphocyte Subsets - cytology
Receptors, Antigen, T-Cell, alpha-beta - metabolism
Staining and Labeling
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Summary:To establish an assay using 9-color flow cytometry immunophenotyping to detect activation and apoptosis of human TCR Vβ lymphocyte subpopulations in peripheral blood samples. We used 5 antibodies (CD3, CD4, CD8, CD95, CD69), phospholipids binding proteins Annexin V, TCR Vβ Repertoire Kit and nucleus dye DAPI to establish a 9-color flow cytometry assay. Peripheral blood samples were taken from eight healthy people for test of antibodies and determination of optimal PMT and staining method (single-stained vs stained with all but one antibody). Appropriate detecting voltage, antibody concentration and compensation methods were determined. The distribution of TCR Vβ subgroup in our samples was consistent with the TCR Vβ Repertoire Kit instruction and other published literature. We have established a effective easy using 9-color flow cytometry immunophenotyping to detect human TCR Vβ lymphocyte subpopulations in peripheral blood samples.
ISSN:1672-173X