TABLE 1.

Optimal parameters of microscopy for mitochondrial swelling measurement with the TR technique

	Important parameters	Additional parameters
Fluorophore	Large molecular weight, organelle-specific, bright	λ is less important, can be a probe
Focus	z-stacking and mean intensity projection	Step size = axial resolution ∼0.8 μm
Optics	NA > 0.8, but higher is better; if confocal, pinhole > 1.5 resel	Wide-field (or confocal)
Detector	0.1–0.14 μm/pixel resolution ∼1000 photo e−/pixel (summed for the z-stack)	Use region averaging