Tracking Motility and Morphology of Individual Mitochondria Reveals Large Heterogeneities in Breast Cancer Subtypes

Mitochondrial dysregulation has long been investigated on the linear microtubules of axons as it makes exploring trafficking relatively simple. In contrast, non-neuronal cells form complex networks which mitochondria can move, and gap in publications mitochondrial dynamics between these cell types illustrates difficulties. Despite this, recent suggest that motion may fuel some metastatic cancer. As exist shapes move erratically across cell, automated segmentation tracking individual are challenging. Consequently, time-consuming manual morphological measurements remain gold standard. These user-dependent methods introduce bias data make them unreliable. Here, we an algorithm for fast, unbiased, spatiotemporal live-cell fluorescence 2D 3D time-lapse images. The inputs pixel size time frames (and 3D, distance stacks) only required from user allow identification changes morphology, motion, fission fusion dynamics. Using a method our lab pioneered assessing metabolic shifts living tissues, phasor-approach to lifetime imaging (FLIM) reduced nicotinamide adenine dinucleotide (NADH), assess cells. We further compare metabolically heterogeneous populations breast lines various potential. Furthermore, using novel technique have developed generating patient-derived xenograft (PDX) spheroids, probe, morphologic, motile, dynamic differences normal epithelia, receptor-positive, triple-negative cancer patients. This work is supported by grant NIH P41-GM103540, NSF GRFP DGE-1839285.