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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 2, Iss. 5 — May. 1, 2011
  • pp: 1030–1039

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  »View Author Affiliations


Biomedical Optics Express, Vol. 2, Issue 5, pp. 1030-1039 (2011)
http://dx.doi.org/10.1364/BOE.2.001030


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Abstract

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 ~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.

© 2011 OSA

OCIS Codes
(170.1470) Medical optics and biotechnology : Blood or tissue constituent monitoring
(180.2520) Microscopy : Fluorescence microscopy
(190.4180) Nonlinear optics : Multiphoton processes
(110.4234) Imaging systems : Multispectral and hyperspectral imaging

ToC Category:
Noninvasive Optical Diagnostics

History
Original Manuscript: February 25, 2011
Revised Manuscript: March 19, 2011
Manuscript Accepted: March 29, 2011
Published: April 1, 2011

Citation
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, "In vivo monitoring of protein-bound and free NADH during ischemia by nonlinear spectral imaging microscopy," Biomed. Opt. Express 2, 1030-1039 (2011)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-2-5-1030


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