Mitochondrial Morphology Imaging in Parasites

Exponentially growing parasites in regular growth medium were collected and resuspended in 10 mL PBS-G (1X PBS, 0.1% glucose). Cells were transferred to flasks and MitoTracker Deep Far Red (ThermoFisher M22426) added at the final concentration of 250 nM. Cells were incubated for 20 minutes at 27°C, collected and fixed with paraformaldehyde and counter-stained with DAPI, and then slides were prepared as described previously [12 (link)]. Slides were visualized on a Carl Zeiss Laser Scanning Microscope 710 using 63x oil DIC objective and MitoTracker excited with the 637nm laser. DAPI was excited with the 405-diode laser. Attained images were analyzed using ZEN 2.3 lite software. Cells were qualitatively compared for the differences in mitochondrion morphology. Representative images from 3 biological replicates are presented.