Traditional fluorescence imaging of oxidative stress and apoptosis using CellROX® Deep Red Reagent (Cat. No. C10422) and CellEvent™ Caspase-3/7 (Cat. No. C10423)
HeLa cells were treated with or without 0.5 µM staurosporine for 2 or 4 hours in the presence of 7.5 µM fluorogenic caspase 3/7 substrate. Cells were then stained with 5 µM Far Red ROS Sensor and Hoechst 33342 for 30 minutes at 37°C, then washed with warm DPBS. Cells were imaged immediately on a Zeiss Axiovert® inverted microscope using a 40x objective. Increased oxidative stress was observed at 2 hours after treatment (magenta) while caspase 3/7 activation was not observed until 4 hours after treatment (green) as shown in representative cells above.