Live Imaging of Oxidative Stress Response

Four days post fertilization (dpf) wildtype larvae were treated with pentafluorobenzenesulfonyl-fluorescein (HPF) and 0.01% (3 mM) H₂O₂ or with HPF alone for 3 hours and static images were recorded using a FV1000 (Olympus) confocal microscope. Uninjured and amputated caudal fins of 3 dpf EPRE:GFP larvae were recorded in 12hr time-lapse movies (one stack every 30 min) on a FV1000 (Olympus) confocal microscope. Three dpf larvae of the Tg(NF-κB:EGFP) reporter strain59 (link) either untreated or treated with 0.01% (3 mM) H₂O₂ were imaged using a FV1000 (Olympus) confocal microscope. Both whole larvae (upper) and tail fins were imaged separately at the beginning and end (2hr post treatment) of an experiment. Quantitative analysis using relative mean fluoresces of the z-stack projected images using ImageJ were performed from three independent experiments using Surface Plot and Measurement tools.