title: Assessment of Cellular Redox State Using NAD(P)H Fluorescence Intensity and Lifetime.
abstract: NADH and NADPH are redox cofactors, primarily involved in catabolic and anabolic metabolic processes respectively. In addition, NADPH plays an important role in cellular antioxidant defence. In live cells and tissues, the intensity of their spectrally-identical autofluorescence, termed NAD(P)H, can be used to probe the mitochondrial redox state, while their distinct enzyme-binding characteristics can be used to separate their relative contributions to the total NAD(P)H intensity using fluorescence lifetime imaging microscopy (FLIM). These protocols allow differences in metabolism to be detected between cell types and altered physiological and pathological states.
authors.affiliation: Consortium for Mitochondrial Research, Department of Cell & Developmental Biology, University College London, London, United Kingdom; Department of Physics & Astronomy, University College London, London, United Kingdom.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Blacker
authors.affiliation: Consortium for Mitochondrial Research, Department of Cell & Developmental Biology, University College London, London, United Kingdom.
authors.forename: Tunde
authors.initials: T
authors.lastname: Berecz
authors.affiliation: Consortium for Mitochondrial Research, Department of Cell & Developmental Biology, University College London, London, United Kingdom.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Duchen
authors.affiliation: Consortium for Mitochondrial Research, Department of Cell & Developmental Biology, University College London, London, United Kingdom; Department of Biomedical Sciences, University of Padua and CNR Neuroscience Institute, Padua, Italy.
authors.forename: Gyorgy
authors.initials: G
authors.lastname: Szabadkai
keywords: Autofluorescence; FLIM; Fluorescence lifetime; Microscopy; NAD(P)H; NADH; NADPH; Redox state