]. In brief, to track lysosomes and mitochondria, cells were double-stained with live cell dyes Lysotracker Red DND-99 and Mitotracker Deep Red FM at a final 50 nM each. Trackers were added from a 1 mM stock in DMSO directly to culture supernatants in both the proximal and distal compartments of MFCs and incubated for 1 h at 37 °C.
, numerical descriptors, for a comprehensive phenotypic profiling of each track through multiple parameterization. Examples of these dynamic tracking parameters comprised organelle mean speed and track displacement; the latter served as a measure for straight, processive movement as opposed to a non-directional random walk. A full list of the obtained parameters is provided below. Likewise, the Morphology macro returned static shape parameters of either type of organelle.