流式细胞术检测细胞内活性氧的方法

Detection of Cellular Reactive Oxygen Species Level by Flow Cytometry

目的流式细胞术检测NB4和U93 7细胞内活性氧 (reactiveoxygenspecies,ROS)的水平。 方法双氢罗丹明 12 3 (DHR)以不同的时间孵育细胞 ( 1、6、2 4h) ,DHR可被细胞内ROS氧化为罗丹明 12 3 ,通过流式细胞仪来检测细胞内罗丹明 12 3的荧光 ,同时设立阴性及阳性对照。 结果 1μmol/LDHR孵育细胞 ,1～ 2 4hNB4和U93 7细胞内的平均荧光强度分别从 19.2 0增加到 3 84 .3 8和 12 .3 1增加到 97.15。 结论适当延长DHR孵育时间可检测不同细胞系ROS基准水平的差异。

Objective To detect the levels of reactive oxygen species (ROS) of NB4 cells and U937 cells by flow cytometry. The fluorescent intensity of cellular oxidized rhodamine 123 was detected by flow cytometry after incubating with dihydrogenrhodamine 123 (DHR) for different periods (1, 6, and 24 hours) and negative controls and positive controls were also set up. After incubation with 1μmol/L DHR from 1 to 24 hours, the mean intensity of oxidized rhodamine 123 increased from 19.20 to 384.38 in NB4 cells, and from 12.31 to 97.15 in U937 cells. The difference of cellular ROS levels can be determined by flow cytometry through prolonging properly the time of incubation with DHR.