Abstract
While fluorescence intensity-based metabolic imaging techniques have provided a useful means of monitoring cellular energetics, recent work has demonstrated that fluorescence lifetime imaging (FLIM) provides additional details into the subcellular trafficking of energy intermediates within the cell. Specifically, FLIM can measure the reorganization of NADH within distinct subcellular pools with change in the metabolic state induced by inhibitors, uncouplers and substrate availability. Here, we compare NADH-intensity and FLIM measurements of metabolism of cells grown as either monolayer culture or 300-500 μm diameter multicell tumor spheroids in media under different growth conditions. These measures are correlated with cellular respiration monitored using an oxygen-sensitive electrodes to test the hypothesis that NADH FLIM-based metabolic imaging more accurately measures the cellular metabolic state of three-dimensional living tissue than intensity-based measurements.This work was supported by P20 RR016469 from the INBRE Program of the National Center for Research Resources, and by NIH R15 GM085776.
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