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Fluorescence lifetime imaging (FLIM) is a powerful tool to quantify local changes in cell metabolism without loss of spatial resolution. Here, we adopted FLIM to characterize spatio-temporal metabolic changes occurring during collective cell migration. Using an established in vitro system, we measured biomechanical and metabolic changes during migration of epithelial cell monolayers. We developed a custom image analysis pipeline that combines machine learning segmentation and curve fitting analysis to analyze FLIM data. Our findings – which were validated via separate measurements of cytoplasmic redox ratio, glucose uptake, and mitochondrial membrane potential – are consistent with a glycolytic shift during collective cell migration.
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Jacopo Ferruzzi, Stephen J. DeCamp, Victor M. K. Tsuda, Muhammad H. Zaman, Jeffrey J. Fredberg, Darren M. Roblyer, "Multiphoton fluorescence lifetime imaging of NADH reveals spatio-temporal patterns in cell metabolism during collective migration," Proc. SPIE 11622, Multiscale Imaging and Spectroscopy II, 116220G (5 March 2021); https://doi.org/10.1117/12.2578743