Research Papers

In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia

[+] Author Affiliations
Melissa C. Skala

Duke University, Department of Biomedical Engineering, Durham, North Carolina 27708

Kristin M. Riching

University of Wisconsin, Department of Biomedical Engineering, Madison, Wisconsin 53706

Damian K. Bird

University of Wisconsin, Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin 53706

Annette Gendron-Fitzpatrick

University of Wisconsin,Research Animal Resources Center, Madison, Wisconsin 53706

Jens Eickhoff

University of Wisconsin,Department of Biostatistics and Medical Informatics, Madison, Wisconsin 53706

Kevin W. Eliceiri

University of Wisconsin,Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin 53706

Patricia J. Keely

University of Wisconsin,Department of Pharmacology, Madison, Wisconsin 53706

Nirmala Ramanujam

Duke University,Department of Biomedical Engineering, Durham, North Carolina 27708

J. Biomed. Opt. 12(2), 024014 (April 10, 2007). doi:10.1117/1.2717503
History: Received July 21, 2006; Revised October 23, 2006; Accepted November 28, 2006; Published April 10, 2007
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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±0.03ns) and long lifetime components (2.03±0.06ns), 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.

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© 2007 Society of Photo-Optical Instrumentation Engineers

Citation

Melissa C. Skala ; Kristin M. Riching ; Damian K. Bird ; Annette Gendron-Fitzpatrick ; Jens Eickhoff, et al.
"In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia", J. Biomed. Opt. 12(2), 024014 (April 10, 2007). ; http://dx.doi.org/10.1117/1.2717503


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