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

Published

Journal Article

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[alpha]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.

Full Text

Cited Authors

  • Skala, MC; Riching, KM; Bird, DK; Gendron-Fitzpatrick, A; Eickhoff, J; Eliceiri, KW; Keely, PJ; Ramanujam, N

Published Date

  • March 2007

Published In

Volume / Issue

  • 12 / 2

Start / End Page

  • 024014 -

PubMed ID

  • 17477729

Pubmed Central ID

  • 17477729

Electronic International Standard Serial Number (EISSN)

  • 1560-2281

International Standard Serial Number (ISSN)

  • 1083-3668

Digital Object Identifier (DOI)

  • 10.1117/1.2717503

Language

  • eng