Share Email Print
cover

Proceedings Paper

Spectroscopic analysis of skin intrinsic signals for multiphoton microscopy
Author(s): Ana-Maria Pena; Mathias Strupler; Thierry Boulesteix; Karim Senni; Gaston Godeau; Emmanuel Beaurepaire; Marie-Claire Schanne-Klein
Format Member Price Non-Member Price
PDF $14.40 $18.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

We recorded multiphoton images of human skin biopsies using endogenous sources of nonlinear optical signals. We detected simultaneously two-photon excited fluorescence (2PEF) from intrinsic fluorophores and second harmonic generation (SHG) from collagen. We observed SHG from fibrillar collagens in the dermis, whereas no SHG was detectable from the non fibrillar type IV collagen in the basal laminae. We compared these distinct behaviours of collagens I and IV in SHG microscopy to polarization-resolved surface SHG experiments on thin films of collagens I and IV molecules. We observed similar signals for both types of molecular films, except for the chiroptical contributions which are present only for collagen I and enhance the signal typically by a factor of 2. We concluded that SHG microscopy is a sensitive probe of the micrometer-scale structural organization of collagen in biological tissues. In order to elucidate the origin of the endogenous fluorescence signals, we recorded 2PEF spectra at various positions in the skin biopsies, and compared these data to in vitro spectroscopic analysis. In particular, we studied the keratin fluorescence and determined its 2PEF action cross section. We observed a good agreement between 2PEF spectra recorded in the keratinized upper layers of the epidermis and in a solution of purified keratin. Finally, to illustrate the capabilities of this technique, we recorded 2PEF/SHG images of skin biopsies obtained from patients of various ages.

Paper Details

Date Published: 22 February 2006
PDF: 7 pages
Proc. SPIE 6078, Photonic Therapeutics and Diagnostics II, 607801 (22 February 2006); doi: 10.1117/12.644777
Show Author Affiliations
Ana-Maria Pena, Lab. for Optics and Biosciences, CNRS, INSERM, Ecole Polytechnique (France)
Mathias Strupler, Lab. for Optics and Biosciences, CNRS, INSERM, Ecole Polytechnique (France)
Thierry Boulesteix, Lab. for Optics and Biosciences, CNRS, INSERM, Ecole Polytechnique (France)
Karim Senni, Paris V Univ. (France)
Gaston Godeau, Paris V Univ. (France)
Emmanuel Beaurepaire, Lab. for Optics and Biosciences, CNRS, INSERM, Ecole Polytechnique (France)
Marie-Claire Schanne-Klein, Lab. for Optics and Biosciences, CNRS, INSERM, Ecole Polytechnique (France)


Published in SPIE Proceedings Vol. 6078:
Photonic Therapeutics and Diagnostics II
Kenton W. Gregory; Nikiforos Kollias; Reza S. Malek; Michael D. Lucroy; Henry Hirschberg; Brian Jet-Fei Wong; Eugene A. Trowers; Werner T.W. de Riese; Justus F. R. Ilgner; Steen J. Madsen; Lloyd P. Tate; Haishan Zeng; Guillermo J. Tearney; Bernard Choi, Editor(s)

© SPIE. Terms of Use
Back to Top