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

作者：Palero Jonathan A^*; Bader Arjen N; de Bruijn Henriette S; van den Heuvel Angelique van der Ploeg; Sterenborg Henricus J C M; Gerritsen Hans C

来源：Biomedical Optics Express, 2011, 2(5): 1030-1039.

Nonlinear spectral imaging microscopy (NSIM) allows simultaneous morphological and spectroscopic investigation of intercellular events within living animals. In this study we used NSIM for in vivo time-lapse in-depth spectral imaging and monitoring of protein-bound and free reduced nicotinamide adenine dinucleotide (NADH) in mouse keratinocytes following total acute ischemia for 3.3 h at similar to 3 min time intervals. The high spectral resolution of NSIM images allows discrimination between the two-photon excited fluorescence emission of protein-bound and free NAD(P) H by applying linear spectral unmixing to the spectral image data. Results reveal the difference in the dynamic response between protein-bound and free NAD(P) H to ischemia-induced hypoxia/anoxia. Our results demonstrate the capability of nonlinear spectral imaging microscopy in unraveling dynamic cellular metabolic events within living animals for long periods of time.

• 出版日期2011-5-1