2',7'-二氯荧光素二乙酸酯是一种荧光探针，作为一种非极性化合物，能够迅速扩散到细胞内，细胞内的酯酶将其水解为非荧光的二氯荧光素（DCFH），从而使探针被装载到细胞内。
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). 1. H2DCFDA is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use. 2. Cells are incubated with 5 µM H2DCFDA solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. 3. Along with the H2DCFDA probe, if indicated, use ROS-insensitive modification of the fluorescein dye DCFDA as a positive control. The staining procedure is the same as for the H2DCFDA.