抗砷细胞荧光染色后的激光共聚焦显微镜检测

Using Laser Scanning Confocal Microscope to Examine Arsenic-resistant Cells after Fluorescence-staining

目的:应用激光共聚焦显微镜检测活细胞内荧光物质含量。方法:传代培养长期低剂量砷诱导的抗砷细胞,用荧光染料Rhodamine-123对细胞染色30min,实验组与维拉帕米(Verapamil)共同孵育,对照组为单加Rhodamine-123的抗砷细胞。应用激光共聚焦显微镜采集Rhodamine-123的荧光图像动态序列,并且记录不同时间段的细胞内荧光强度。结果:实验组细胞染色12h,24h,36h,48h,60h后,荧光强度依次为(51.567±0.7572)、(46.533±0.7095)、(39.557±0.601)、(38.6±0.6245)和(38.505±0.718),明显高于同时间段对照组的荧光强度,差异均有显著性(P<0.01)。结论:应用激光共聚焦显微成像技术能进行活细胞水平荧光物质实时定量检测。

Objective： To examine the fluorescent material by laser scanning confocal microscope （LSCM） in living cells. Methods： The arsenic-resistant cells were incubated with fluorescent dyes Rhodamine-123 for 30 minutes. The test group was co-incubated with Verapamil, and the arsenic-resistant cells were merely administrated with Rhodamine-123 as control group. Fluorescence image acquisition of Rhodamine-123＇s dynamic series was executed by the LSCM, and the intracellular fluorescence intensity were recorded during different times. Results： After fluorescence staining for 12, 24, 36, 48, 60h, the fluorescence intensity of test group were 51.567±0.7572, 46.533±0.7095, 39.557±0.601, 38.6±0,6245 and 38.505±0.718 orderly. They were significantly higher than those of the synchronous control group, with statistically significant differences （p〈0.01）. Conclusions： The LSCM could be used for content detection of fluorescence in living cells dynamically and quantitively.