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Characterizing Population-level Heterogeneities in Mammalian Organelle Morphology

From Q-bio

Organelles are specialized cellular compartments responsible for carrying out a wide range of biochemical processes necessary for cellular growth and survival. Cellular organelles are characterized by well-defined morphologies, yet the functional importance of these morphologies remains incompletely understood. To better understand the role organelle morphology plays in cellular function, we are conducting experiments to quantify the native distribution of organelle morphology in WT mammalian cells. By characterizing heterogeneity in organelle morphology, we will be able to identify morphological subpopulations and investigate whether and how morphological heterogeneity coincides with variability in cellular behavior. This variability may include changes in transcription levels, epigenetic modifications, intracellular trafficking, and ATP production, amongst others. We have developed a high-content imaging assay to quantify morphological heterogeneity. We have labeled a genetically clonal population of mouse embryonic fibroblasts with fluorescent markers for the nucleus, mitochondria, and cytoskeleton. We follow with automated, high-throughput imaging, image processing, and parameterization of the organelles/structures of interest. Eventual statistical analysis will provide insight into native variability in organelle morphology and lay the foundation for future studies of the relationship between organelle morphologies, cellular behavior, and disease states.