Confocal Microscopy and Image Analysis
All slides were imaged on a Nikon A1R confocal microscope using either a 40X or 60X objective. To analyze the percent area occupied by a specific organelle, 10 fields per slide were imaged using the 40X objective. These frames were processed using an in-house plugin for FIJI (National Institutes of Health), which contained components inspired by the ImageJ Mitochondrial Morphology Macro (Ruben K. Dagda Ph.D.; University of Nevada School of Medicine, Stefan Strack Ph.D.; University of Iowa, and Charleen Chu; University of Pittsburgh). Briefly, cell perimeters were manually outlined, the channels were split, thresholded, and analyzed for percent area of the appropriate channel.