Comparison of MitoTracker and a new mitochondrial marker for fluorescence imaging. (A) Fluorescent imaging of WT parasites stained with MitoTracker Orange CMTMRos (MT orange) or MitoTracker Red CMXRos (MT red). (B) Fluorescence microscopy of MitoRed. The mito-mScarlet signal was observed in all asexual life-cycle stages including rings, trophozoites, and early and late schizonts. No antibody staining was used and the fluorescent signal observed is exclusively the mito-mScarlet signal. (C) Fluorescence microscopy of MitoRed, either unstained (no MT) or stained with MT orange, MT red, or MitoTracker Deep Red FM (MT deep red). Mito signal is the combined MitoTracker and mito-mScarlet signal that is observed in this channel. DAPI (blue) is used to visualize DNA and DIC (differential interference contrast) for general cellular context. All images are maximum intensity projections of Z-stacks (41 slices, 150 nm interval) taken with an Airyscan confocal microscope. Scale bars, 2 μm.