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Proceedings Paper

Fluorescence spectroscopy of biological tissue: single- and two-photon excitation
Author(s): Yicong Wu; Peng Xi; Weikun Ge; Powing Yuen; Jianan Y. Qu
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Paper Abstract

Endogenous fluorophores, such as NAD(P)H/FAD and collagen/elastin, have been regarded as in vivo quantitative fluorescence biomarkers for precancerous changes of epithelial tissue. However, the fluorescence signal measured by conventional spectroscopy is a mixture of autofluorescence from the epithelium and deep structures. The dominant fluorescence of collagen/elastin from connective tissue in deep layers creates serious challenge for extracting the epithelial fluorescence of NAD(P)H/FAD that is weak, but important for the characterization of tissue pathology. In this work, we instrumented a confocal fluorescence spectroscopy system and a two-photon excited fluorescence spectroscopy system to measure the depth-resolved single- and two-photon fluorescence spectra from the rabbit esophageal tissues. The excitation wavelengths were 349 nm and 735 nm, respectively. Both systems provided good optical sectioning. The information obtained from depth-resolved fluorescence was generally consistent with the histology of the examined tissue sample. The NAD(P)H signals from epithelial layers were clearly separated from the collagen signal from deep layers. In addition, strong second harmonic generations given by collagen fibers were observed. This work demonstrates that depth-resolved fluorescence spectroscopy may produce more accurate information on the diagnosis of tissue pathology.

Paper Details

Date Published: 21 June 2004
PDF: 5 pages
Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); doi: 10.1117/12.544404
Show Author Affiliations
Yicong Wu, Hong Kong Univ. of Science and Technology (Hong Kong China)
Peng Xi, Hong Kong Univ. of Science and Technology (Hong Kong China)
Weikun Ge, Hong Kong Univ. of Science and Technology (Hong Kong China)
Powing Yuen, Univ. of Hong Kong (Hong Kong China)
Jianan Y. Qu, Hong Kong Univ. of Science and Technology (Hong Kong China)

Published in SPIE Proceedings Vol. 5323:
Multiphoton Microscopy in the Biomedical Sciences IV
Ammasi Periasamy; Peter T. C. So, Editor(s)

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