1 March 2007 In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia
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J. of Biomedical Optics, 12(2), 024014 (2007). doi:10.1117/1.2717503
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.
Melissa C. Skala, Kristin M. Riching, Damian K. Bird, Annette Gendron-Fitzpatrick, Jens Eickhoff, Kevin W. Eliceiri, Patricia J. Keely, Nirmala Ramanujam, "In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia," Journal of Biomedical Optics 12(2), 024014 (1 March 2007). http://dx.doi.org/10.1117/1.2717503
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KEYWORDS
In vivo imaging

Fluorescence lifetime imaging

Luminescence

Tissues

Multiphoton fluorescence microscopy

Mode conditioning cables

Tumor growth modeling

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