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Non-linear microscopy and spectroscopy of skin tissues
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Paper Abstract

We combined a non-linear microscope with a sensitive prism-based spectrograph and employed it for the imaging of the auto fluorescence of skin tissues. The system has a sub-micron spatial resolution and a spectral resolution of better than 5 nm. The spectral images contain signals arising from two-photon excited fluorescence (TPEF) of endogenous fluorophores in the skin and from second harmonic generation (SHG) produced by the collagen fibers, which have non-centrosymmetric structure. Non-linear microscopy has the potential to image deep into optically thick specimens because it uses near-infrared (NIR) laser excitation. In addition, the phototoxicity of the technique is comparatively low. Here, the technique is used for the spectral imaging of unstained skin tissue sections. We were able to image weak cellular autofluorescence as well as strong collagen SHG. The images were analyzed by spectral unmixing and the results exhibit a clear spectral signature for the different skin layers.

Paper Details

Date Published: 25 October 2005
PDF: 9 pages
Proc. SPIE 5968, Laser Florence 2004: A Window on the Laser Medicine World, 59680H (25 October 2005); doi: 10.1117/12.660069
Show Author Affiliations
Jonathan A. Palero, Utrecht Univ. (Netherlands)
Gwendal Latouche, LURE/CNRS, Ctr. Univ. Paris sud (France)
Henriëtte S. de Bruijn, Univ. Medical Ctr. Rotterdam (Netherlands)
Hans C. Gerritsen, Univ. Utrecht (Netherlands)
Henricus J. C. M. Sterenborg, Univ. Medical Ctr. Rotterdam (Netherlands)

Published in SPIE Proceedings Vol. 5968:
Laser Florence 2004: A Window on the Laser Medicine World
Leonardo Longo; Khalil A. Khatri; Mihail-Lucian Pascu; Wilhelm R.A. Waidelich, Editor(s)

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