Metabolic control of immune cell function has emerged as a key regulatory mechanism in immunity. Current studies rely mainly on in vitro and ex vivo approaches to characterize the intracellular metabolism of immune cells. Here, we show that fluorescence lifetime imaging microscopy of metabolic coenzymes, NAD(P)H and FAD, captures dynamic changes in the metabolic activity of macrophages during diverse inflammatory responses in their native interstitial microenvironments. This imaging-based approach also provides single-cell resolution, in contrast to bulk measurements provided by traditional biochemical assays, making it a potentially valuable tool in the field of immunometabolism.
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