流式细胞术_____钙离子；Fluo3

双色流式细胞术在检测特定亚群淋巴细胞Ca2+变化中的应用

http://www.100md.com 《第四军医大学学报》 2004年第4期

流式细胞术,,钙离子；Fluo3,AM；流式细胞术,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Application of doublecolor flow cytometry in detecting intracellular Ca2+ mobilization of certain lymphocytic subsets

    CAO YunXin, HAN WeiNing, JIN BoQuan

    Department of Immunology, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To establish a method of detecting intracellular Ca2+ mobilization of certain lymphocytic subsets. METHODS: Ninetysix hours after PBMC were stimulated by PHA, the cell surface was stained with quantumredlabeled antiCD8 mAb and loaded with Fluo3/AM. The intracellular Ca2+ mobilization of CD8+ lymphocytes was examined by flow cytometry to find an optimal condition for doublecolor flow cytometry analysis of the intracellular Ca2+ mobilization. RESULTS: ① 1 mol/L was decided upon as the optimal Fluo3/AM loading concentration; ② The modified cell surface immunofluorescence staining condition corresponding to the Fluo3/AM loading did not affect the positive proportion and mean fluorescence intensity; ③ The intracellular Ca2+ loading level increased after chemokinic stimulation, which, however, was far less significant than that of A23187induced Ca2+ mobilization. CONCLUSION: A method for a quick and accurate detection of intracellular Ca2+ mobilization of certain lymphocytic subsets by doublecolor flow cytometry analysis has been established. ......

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