Visualizing Mitochondria in C. elegans

Whole animal images were acquired on an Olympus MVX10 Fluorescence MacroZoom dissecting microscope, using a 540-580 nm excitation filter and 590-670 nm emission filter. Mitochondrial images were taken on a FV1000 Olympus laser scanning confocal microscope using a 100x oil objective (Olympus, N.A. 1.40). Diode laser illumination was 561 nm for red fluorescent transgene and 488 nm for fluorescence of MitoTracker Green FM. Animals were stained with 12 μM MitoTracker Green FM for 20 hr where indicated. MitoTracker stain was dissolved in DMSO, diluted in M9 media (22 mM KH₂PO₄, 42 mM Na₂HPO₄, 86 mM NaCl, 1 mM MgSO₄, pH 7) and added to OP50 food on culture plates (DMSO <0.02% final concentration) and allowed to dry (Dingley et al. 2010 (link)). Profile plots of pixel intensity were generated using ImageJ software.