Subsequent to the indicated drug treatment for 24 h, the supernatant was discarded. The cells were rinsed with PBS, and stained with 10 µM DCFH-DA solution in the dark for 30 min in accordance with the reactive oxygen species assay kit (cat. no. CA1410; Beijing Solarbio Science & Technology Co., Ltd.) protocol (28 (link)). Inverted fluorescence microscopy (DMI3000, Leica) was used to record the morphological changes. The stained cells were analyzed by FACSCanto II flow cytometer (BD Biosciences). The fluorescence mean value in P2 from the flow cytometer was collected using FACSDiva software (version 6.1.3, BD Biosciences), which was used for quantitative analysis. The data were presented as fold increase normalized to the control group.
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Quantifying Intracellular Reactive Oxygen
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Corresponding Organization :
Other organizations : Ocean University of China, Binzhou Medical University, Binzhou University, Nantong University
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Variable analysis
independent variables
- Drug treatment
- Reactive oxygen species (ROS) levels
- Morphological changes
- Incubation time (24 h)
- Staining with DCFH-DA solution (10 µM, 30 min)
- Inverted fluorescence microscopy
- Flow cytometry analysis (FACSCanto II)
- Control group
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