Correlative NAD(P)H‐FLIM and oxygen sensing‐PLIM for metabolic mapping. Issue 8 (15th March 2016)

• Record Type: Journal Article
• Authors: Kalinina, Sviatlana; Breymayer, Jasmin; Schäfer, Patrick; Calzia, Enrico; Shcheslavskiy, Vladislav; Becker, Wolfgang; Rück, Angelika
• Other Names: Vitkin Alex guestEditor.; Kirillin Mikhail guestEditor.; Zagaynova Elena guestEditor.
• Is Part Of: Journal of biophotonics. Volume 9:Issue 8(2016)
• Page Start: 800
• Page End: 811
• Publication Date: 2016-03-15
• Subjects: FLIM -- PLIM -- NAD(P)H -- redox state -- oxygen tension -- cell metabolism -- respiratory chain -- antimycin A; Photonics -- Periodicals; Optical materials -- Periodicals; Optics -- Periodicals; Medical instruments and apparatus -- Periodicals; 621.3605
• DOI: 10.1002/jbio.201500297
• Languages: English
• ISSNs: 1864-063X

Abstract

Abstract : Cellular responses to oxygen tension have been studied extensively. Oxygen tension can be determined by considering the phosphorescence lifetime of a phosphorescence sensor. The simultaneous usage of FLIM of coenzymes as NAD(P)H and FAD + and PLIM of oxygen sensors could provide information about correlation of metabolic pathways and oxygen tension. We investigated correlative NAD(P)H‐FLIM and oxygen sensing‐PLIM for simultaneously analyzing cell metabolism and oxygen tension. Cell metabolism and p O2 were observed under different hypoxic conditions in squamous carcinoma cell cultures and in complex ex vivo systems. Increased hypoxia induced an increase of the phosphorescence lifetime of Ru(BPY)3 and in most cases a decrease in the lifetime of NAD(P)H which is in agreement to the expected decrease of the protein‐bound NAD(P)H during hypoxia. Oxygen was modulated directly in the mitochondrial membrane. Blocking of complex III and accumulation of oxygen could be observed by both the decrease of the phosphorescence lifetime of Ru(BPY)3 and a reduction of the lifetime of NAD(P)H which was a clear indication of acute changes in the redox state of the cells. For the first time simultaneous FLIM/PLIM has been shown to be able to visualize intracellular oxygen tension together with a change from oxidative to glycolytic phenotype. Abstract : The simultaneous usage of FLIM of coenzymes as NAD(P)H and PLIM of oxygen sensors could provide information about correlation of metabolic pathways and oxygen tension in cells and tissue. Investigation of correlative NAD(P)H‐FLIM and oxygen sensing‐PLIM has been shown to be able to visualize variations in intracellular oxygen tension together with a change of metabolic phenotypes.