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Optics Letters

Optics Letters


  • Editor: Alan E. Willner
  • Vol. 34, Iss. 2 — Jan. 15, 2009
  • pp: 202–204

Two-photon autofluorescence microscopy of multicolor excitation

Dong Li, Wei Zheng, and Jianan Y. Qu  »View Author Affiliations

Optics Letters, Vol. 34, Issue 2, pp. 202-204 (2009)

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We developed a two-photon autofluorescence lifetime imaging system with excitations selected from the supercontinuum generated from a photonic crystal fiber. The system excites multiple endogenous fluorophores, such as nicotinamide adenine dinucleotide (NADH) and tryptophan, simultaneously and produces coregistered two-photon autofluorescence images of a biological sample. The technology provides a unique approach to investigate the cellular metabolic activity and protein expression in cells that are potentially important for noninvasive precancer diagnostics. We demonstrated that by taking the tryptophan fluorescence as a reference the ratio of NADH to the tryptophan signal serves as a sensitive indicator of cellular metabolism. The ratio can also clearly differentiate normal cells from cancer cells. The tryptophan fluorescence lifetime images of cells shows that the lifetime of tryptophan fluorescence, varying over a wide range, may be highly dependent on the expression and structure of the protein that tryptophan is packed in.

© 2009 Optical Society of America

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(300.6410) Spectroscopy : Spectroscopy, multiphoton
(300.6500) Spectroscopy : Spectroscopy, time-resolved
(180.4315) Microscopy : Nonlinear microscopy
(320.6629) Ultrafast optics : Supercontinuum generation

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: September 29, 2008
Revised Manuscript: November 18, 2008
Manuscript Accepted: November 22, 2008
Published: January 14, 2009

Virtual Issues
Vol. 4, Iss. 3 Virtual Journal for Biomedical Optics

Dong Li, Wei Zheng, and Jianan Y. Qu, "Two-photon autofluorescence microscopy of multicolor excitation," Opt. Lett. 34, 202-204 (2009)

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