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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 1, Iss. 6 — Jun. 13, 2006

Time-resolved optical imaging provides a molecular snapshot of altered metabolic function in living human cancer cell models

Dhruv Sud, Wei Zhong, David G. Beer, and Mary-Ann Mycek  »View Author Affiliations

Optics Express, Vol. 14, Issue 10, pp. 4412-4426 (2006)

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A fluorescence lifetime imaging microscopy (FLIM) method was developed and applied to investigate metabolic function in living human normal esophageal (HET-1) and Barrett’s adenocarcinoma (SEG-1) cells. In FLIM, image contrast is based on fluorophore excited state lifetimes, which reflect local biochemistry and molecular activity. Unique FLIM system attributes, including variable ultrafast time gating (≥200 ps), wide spectral tunability (337.1–960 nm), large temporal dynamic range (≥600 ps), and short data acquisition and processing times (15 s), enabled the study of two key molecules consumed at the termini of the oxidative phosphorylation pathway, NADH and oxygen, in living cells under controlled and calibrated environmental conditions. NADH is an endogenous cellular fluorophore detectable in living human tissues that has been shown to be a quantitative biomarker of dysplasia in the esophagus. Lifetime calibration of an oxygen-sensitive, ruthenium-based cellular stain enabled in vivo oxygen level measurements with a resolution of 8 µM over the entire physiological range (1–300 µM). Starkly higher intracellular oxygen and NADH levels in living SEG-1 vs. HET-1 cells were detected by FLIM and attributed to altered metabolic pathways in malignant cells.

© 2006 Optical Society of America

OCIS Codes
(170.1530) Medical optics and biotechnology : Cell analysis
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(170.4580) Medical optics and biotechnology : Optical diagnostics for medicine
(170.6920) Medical optics and biotechnology : Time-resolved imaging

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: February 14, 2006
Revised Manuscript: May 8, 2006
Manuscript Accepted: May 8, 2006
Published: May 15, 2006

Virtual Issues
Vol. 1, Iss. 6 Virtual Journal for Biomedical Optics

Dhruv Sud, Wei Zhong, David G. Beer, and Mary-Ann Mycek, "Time-resolved optical imaging provides a molecular snapshot of altered metabolic function in living human cancer cell models," Opt. Express 14, 4412-4426 (2006)

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