应用流式细胞分析仪同时分析细胞表面抗原及DNA含量

SIMULTANEOUS ANALYSIS OF CELL SURFACE MARKER AND DNA CONTENT

作者在应用流式细胞分析仪用于分析 DNA 和细胞表面标记的基础上,进一步发展成同时分析 DNA 和细胞表面抗原。其法是首先将细胞用结合荧光的抗体染色,在免疫染色后,再用0.5%多聚甲醛100μl 将细胞固定5分钟,然后以碘化丙啶(Propidium iodide,PI)染色,避光保存。用0.5%多聚甲醛固定能很好的保存细胞表面结合韵抗体,经 DNA 分析,变异系数(CV)平均为4%(2.1%～6.3%)。在细胞表面抗原呈现的基础上可鉴别细胞的类型,同时通过细胞表面抗原与细胞周期相结合的方法进行分析。

Flow cytometry is a useful tool for measuring DNA content and analysis of cell sur-face marker.We have developed the technigue of simultaneous analysis of cell surface anti-gen and DNA content.Cells are first stained with fluorescein isothiocyanate conjugated(FI- TC)antibody,then the cells were fixed by 0.5% paraformaldehyde 100 nl,and 5 minlater the cells were stained with propidium iodide in darkness at 4℃.Fixation with 0.5%paraformaldehyde could preserve cell surface binding antibodies.The coefficient of vari-ation for DNA content was 2.1-6.3%,average 4%.It is possible to identify cell typeby combination of cell surface antigen expression and the cell cycle.