2.8. Measurement of Intracellular ROS Production in BV2 Microglia

BV2 microglia were cultured at a density of 5 × 10⁴ cells/well in 24-well plates. After the corresponding treatments were administered, intracellular ROS production was measured in BV2 microglia using the DCFH-DA fluorescent probe (Beyotime). The microglia were incubated with 10 μM DCFH-DA for 30 min in the dark at 37°C. Fluorescence images were observed under an inverted fluorescence microscope (DMI4000B, Leica, Germany). The fluorescence intensity of images was analyzed by ImageJ software (BX50-FLA, Olympus), and results are expressed as percentages of control levels.