Time-resolved spectroscopic imaging reveals the fundamentals of cellular NADH fluorescence

Abstract

A time-resolved spectroscopic imaging system is built to study the fluorescence characteristics of nicotinamide adenine dinucleotide (NADH), an important metabolic coenzyme and endogenous fluorophore in cells. The system provides a unique approach to measure fluorescence signals in different cellular organelles and cytoplasm. The ratios of free over protein-bound NADH signals in cytosol and nucleus are slightly higher than those in mitochondria. The mitochondrial fluorescence contributes about 70% of overall cellular fluorescence and is not a completely dominant signal. Furthermore, NADH signals in mitochondria, cytosol, and the nucleus respond to the changes of cellular activity differently, suggesting that cytosolic and nuclear fluorescence may complicate the well-known relationship between mitochondrial fluorescence and cellular metabolism.

© 2008 Optical Society of America

In vivo monitoring of protein-bound and free NADH during ischemia by nonlinear spectral imaging microscopy

Jonathan A. Palero, Arjen N. Bader, Henriëtte S. de Bruijn, Angélique van der Ploeg van den Heuvel, Henricus J. C. M. Sterenborg, and Hans C. Gerritsen
Biomed. Opt. Express 2(5) 1030-1039 (2011)

Sensing cell metabolism by time-resolved autofluorescence

Yicong Wu, Wei Zheng, and Jianan Y. Qu
Opt. Lett. 31(21) 3122-3124 (2006)

Two-photon autofluorescence microscopy of multicolor excitation

Dong Li, Wei Zheng, and Jianan Y. Qu
Opt. Lett. 34(2) 202-204 (2009)

Fluorescence quenching of free and bound NADH in HeLa cells determined by hyperspectral imaging and unmixing of cell autofluorescence

Aziz Ul Rehman, Ayad G. Anwer, Martin E. Gosnell, Saabah B. Mahbub, Guozhen Liu, and Ewa M. Goldys
Biomed. Opt. Express 8(3) 1488-1498 (2017)

Fluorescence lifetime microscopy of NADH distinguishes alterations in cerebral metabolism in vivo

Mohammad A. Yaseen, Jason Sutin, Weicheng Wu, Buyin Fu, Hana Uhlirova, Anna Devor, David A. Boas, and Sava Sakadžić
Biomed. Opt. Express 8(5) 2368-2385 (2017)