The dynamic nature of the mitochondria varies with cellular needs, developmental stage, and tissue type, with the maintenance of structural integrity a necessary feature for mitochondria to function effectively. The general morphology of the mitochondria is organized into an elaborate network that dynamically interconverts through frequent rounds of fission and fusion events that are tightly regulated to control overall morphology ( 4– 8). The functioning of the mitochondria is intimately linked to dramatic modifications to its structure and remodeling that occurs within ( 2– 5). Mitochondria are essential organelles responsible for a host of cellular functions, with a primary objective of meeting the cellular bioenergetics demands via the production of adenosine triphosphate (ATP) ( 1). Our results demonstrate that MitoCellPhe enables high-throughput, sensitive, detailed, and quantitative mitochondrial morphological assessment and thus enables better biological insights into mitochondrial dynamics in health and disease. Using images generated from control and diseased fibroblasts and hiPSCs, we also demonstrate the efficacy of the toolset in delineating differences in morphologies between healthy and the diseased state in both stem cell (hiPSC) and differentiated fibroblast cells. With this tool, we were able to validate previous findings that show networks of mitochondria in healthy fibroblast cell lines and a more fragmented morphology in hiPSCs. MitoCellPhe generates 24 parameters, allowing for a comprehensive analysis of mitochondrial structures and importantly allows for quantification to be performed on mitochondria in images containing single cells or clusters of cells. Here, we present the Mitochondrial Cellular Phenotype (MitoCellPhe) tool developed for quantifying mitochondrial morphologies and demonstrate its utility in delineating differences in mitochondrial morphologies in a human fibroblast and human induced pluripotent stem cell (hiPSC) line. Despite the wide biological significance in disease and in health, delineating mitochondrial morphologies in complex systems remains challenging. Mitochondria are dynamic organelles that differ significantly in their morphologies across cell types, reflecting specific cellular needs and stages in development.
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