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Autofluorescence lifetime imaging is a useful tool to quantify features of cellular metabolism. Here, we use multiphoton fluorescence lifetime imaging to measure NADH and FAD fluorescence lifetimes. We compared fluorescence intensity and lifetime features of T cells treated with a panel of metabolic inhibitors to correlate imaging features with metabolic pathways. Differences between autofluorescence features of T cells and cancer cells allow robust classification of cell type within simulations of complex tumor tissues. Autofluorescence lifetime imaging combined with automated image segmentation, analysis, and classification enables robust and label-free determination of cell type and function.
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Alex J. Walsh, Linghao Hu, Elizabeth Cardona, Nianchao Wang, Jocelyn Martinez, Uyen Nguyen, Naman Bala, "Fluorescence lifetime imaging of metabolic co-enzymes to determine cell type and function," Proc. SPIE 11648, Multiphoton Microscopy in the Biomedical Sciences XXI, 116480K (5 March 2021); https://doi.org/10.1117/12.2577866