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Journal of Biomedical Optics

In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia
Author(s): Melissa C. Skala; Kristin M. Riching; Damian K. Bird; Annette Gendron-Fitzpatrick; Jens Eickhoff; Kevin W. Eliceiri; Patricia J. Keely; Nirmala Ramanujam
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Paper Abstract

Multiphoton fluorescence lifetime imaging microscopy (FLIM) is a noninvasive, cellular resolution, 3-D functional imaging technique. We investigate the potential for in vivo precancer diagnosis with metabolic imaging via multiphoton FLIM of the endogenous metabolic cofactor nicotinamide adenine dinucleotide (NADH). The dimethylbenz[α]anthracene (DMBA)-treated hamster cheek pouch model of oral carcinogenesis and MCF10A cell monolayers are imaged using multiphoton FLIM at 780-nm excitation. The cytoplasm of normal hamster cheek pouch epithelial cells has short (0.29±0.03 ns) and long lifetime components (2.03±0.06 ns), attributed to free and protein-bound NADH, respectively. Low-grade precancers (mild to moderate dysplasia) and high-grade precancers (severe dysplasia and carcinoma in situ) are discriminated from normal tissues by their decreased protein-bound NADH lifetime (p<0.05). Inhibition of cellular glycolysis and oxidative phosphorylation in cell monolayers produces an increase and decrease, respectively, in the protein-bound NADH lifetime (p<0.05). Results indicate that the decrease in protein-bound NADH lifetime with dysplasia is due to a shift from oxidative phosphorylation to glycolysis, consistent with the predictions of neoplastic metabolism. We demonstrate that multiphoton FLIM is a powerful tool for the noninvasive characterization and detection of epithelial precancers in vivo.

Paper Details

Date Published: 1 March 2007
PDF: 10 pages
J. Biomed. Opt. 12(2) 024014 doi: 10.1117/1.2717503
Published in: Journal of Biomedical Optics Volume 12, Issue 2
Show Author Affiliations
Melissa C. Skala, Duke Univ. (United States)
Kristin M. Riching, Univ. of Wisconsin/Madison (United States)
Damian K. Bird, The Univ. of Melbourne (Australia)
Annette Gendron-Fitzpatrick, Univ. of Wisconsin/Madison (United States)
Jens Eickhoff, Univ. of Wisconsin/Madison (United States)
Kevin W. Eliceiri, Univ. of Wisconsin/Madison (United States)
Patricia J. Keely, Univ. of Wisconsin/Madison (United States)
Nirmala Ramanujam, Duke Univ. (United States)

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