粘附式细胞仪测试高温对培养巨噬细胞DNA荧光强度的影响

Analysis and test of fluorescence intensity of heat-treated mouse peritoneal cavity macrophages with adherent cell and sorting cytometer

将体外培养４－５天的小鼠腹腔巨噬细胞分成４组，分别置于３７℃、３９℃、４１℃、４３℃恒温环境再培养２ｈ。细胞用４，６－二脒基－２－苯基吲哚（ＤＡＰＩ）染色后，用ＡＣＡＳ５７０激光细胞仪进行测试和分析。以综合荧光量和平均荧光量作为判断高温对细胞中ＤＮＡ荧光强度影响的主要参数，结果显示，３９℃组细胞中ＤＮＡ荧光强度降低，但４１℃、４３℃组的又缓慢上升，形成弧形曲线。

unming mouse peritoneal cavity macrophages were cultured for 4-5 days in vitro. The cells were then divided into 4 groups,and incubated at 37 ℃,39 ℃, 4l℃and 43℃for 2h respectivly. The fluorescence intensity of the macrophages was analysed and testedwith adherent cell analysis and sorting cytometre after staining with the DNA-specific dye 4,6-diamidino-2-phenylindole(DAPI).The integrated and average fluorescence values were taken as the main parameters of DNA fluorescence intensity changes of heat-treated cells,The results suggested that the fluorescence intensity of 39℃-treated cells was lower as compared with the control(37℃ ),but the fluorescence intensity of 41℃- treated cells rose slowly again,and, therefore,formed a curve with the shape of an inverted saddle.