I am planning a live-cell imaging experiment to study mitochondrial dynamics in cultured HeLa cells. The mitochondria are labeled with MitoTracker Red like Scarlet App, and my goal is to track changes in mitochondrial morphology over time under drug treatment.
I plan to acquire time-lapse images over several hours, but I am concerned about phototoxicity and photobleaching, especially since mitochondrial function is sensitive to light exposure. The cells are grown on glass-bottom dishes, and I have access to widefield and confocal microscopes.
I would appreciate advice on which imaging modality would be most appropriate for this experiment and how to optimize acquisition settings to minimize phototoxic effects while still obtaining sufficient spatial and temporal resolution. In particular, I am interested in recommendations regarding laser power, exposure time, and imaging intervals.