Intracellular Reactive Oxygen Species (ROS) Detection

The cell-permeable dye DCFH-DA was used to detect ROS production, whereby the ROS could oxidize the non-fluorescent dye to green fluorescent of DCF. In a typical setup, 2×10⁵ HepG2 cells were seeded and incubated overnight. Subsequently, it was treated with LP@MnAs_x for 6 h. Then, the culture medium was then replaced with DCFH-DA (10 µM), and then incubated at 37°C for 30 min. Finally, the sample was observed under a confocal laser scanning microscope (CLSM, Olympus, Tokyo, Japan). And the quantitative analysis of DCF with different treatments was assessed via flow cytometry (Guava Easycyte, Merck Millipore, Darmstadt, Germany).