Huang Qiao-rong, Wen Bo, Li Xue, Hu Lian, Wan Xue-mei, Zeng Sheng-ping, Meng Wen-tong
Sichuan Da Xue Xue Bao Yi Xue Ban. 2016 Mar;47(2):262-6.
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.
建立一种使用九色流式细胞术免疫表型分析来检测外周血样本中人类TCR Vβ淋巴细胞亚群的活化和凋亡的方法。
我们使用5种抗体（CD3、CD4、CD8、CD95、CD69）、磷脂结合蛋白膜联蛋白V、TCR Vβ谱试剂盒和细胞核染料DAPI建立了一种九色流式细胞术检测方法。采集8名健康人的外周血样本进行抗体检测以及最佳光电倍增管（PMT）和染色方法（单染与除一种抗体外的所有抗体染色）的确定。
确定了合适的检测电压、抗体浓度和补偿方法。我们样本中TCR Vβ亚群的分布与TCR Vβ谱试剂盒说明书及其他已发表文献一致。
我们建立了一种有效的、易于使用的九色流式细胞术免疫表型分析方法来检测外周血样本中的人类TCR Vβ淋巴细胞亚群。
