The real‐time quantification of autofluorescence spectrum shape for the monitoring of mitochondrial metabolism

The cellular proportion of free and protein‐bound NADH complexes is increasingly recognized as a metabolic indicator and biomarker. Because free and bound forms exhibit different fluorescence spectra, we consider whether autofluorescence shape sufficiently correlates with mitochondrial metabolism to be useful for monitoring in cellular suspensions. Several computational approaches for rapidly quantifying spectrum shape are used to detect Saccharomyces cereviseae response to oxygenation, and to the addition of mitochondrial functional modifiers and metabolic substrates. Observed changes appear consistent with previous studies probing free/protein‐bound proportions, making this a potentially useful approach for the real‐time monitoring of metabolism. (© 2015 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)