Quantify Mitochondrial Dynamics in Cells

To quantify donor mitochondria internalization, fusion with host mitochondria, and mitophagy of donor and host mitochondria (per colocalization with lysosomes), stained CM were co-incubated with pHrodo Red-stained donor mitochondria. 3–10 cells/dish were imaged at baseline and after 1, 4, and 18 h co-incubation using an Eclipse A1R microscope and NIS-Elements software (Nikon Instruments Inc., Melville, NY, USA). Images were analyzed as previously described [9 (link)] using High-Content Screening Navigator (HCS) Colocalization.V4 protocol (Perkin Elmer, Waltham, MA, USA); non-internalized donor mitochondria were excluded. Baseline characteristics were evaluated using HCS Morphology.V4 protocol.