2.9. DCFH-DA assay

To detect ROS generation, 2, 7-dichlorofluorescein diacetate (DCFH-DA) assay was employed. Basically, cells (1 × 10⁶/well) were seeded in 6-well culture plates. Following overnight culture, the cells were treated with AgNPs for 24 and 48 hr time interval. After these, the cells were harvested and re-suspended in PBS. Following this, the cells were incubated with DCFH-DA reagent for 30 min at RT. The samples were then analyzed with a flow cytometer (MACSQuant, Germany) following standard flow procedures. For fluorescence microscopic analysis, the cells were analysed under fluorescence microscope.