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Proceedings Paper

Nonlinear microscopy of collagen fibers
Author(s): M. Strupler; A.-M. Pena; M. Hernest; P.-L. Tharaux; A. Fabre; J. Marchal-Somme; B. Crestani; D. Débarre; J.-L. Martin; E. Beaurepaire; M.-C. Schanne-Klein
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Paper Abstract

We used intrinsic Second Harmonic Generation (SHG) by fibrillar collagen to visualize the three-dimensional architecture of collagen fibrosis at the micrometer scale using laser scanning nonlinear microscopy. We showed that SHG signals are highly specific to fibrillar collagen and provide a sensitive probe of the micrometer-scale structural organization of collagen in tissues. Moreover, recording simultaneously other nonlinear optical signals in a multimodal setup, we visualized the tissue morphology using Two-Photon Excited Fluorescence (2PEF) signals from endogenous chromophores such as NADH or elastin. We then compared different methods to determine accurate indexes of collagen fibrosis using nonlinear microscopy, given that most collagen fibrils are smaller than the microscope resolution and that second harmonic generation is a coherent process. In order to define a robust method to process our three-dimensional images, we either calculated the fraction of the images occupied by a significant SHG signal, or averaged SHG signal intensities. We showed that these scores provide an estimation of the extension of renal and pulmonary fibrosis in murine models, and that they clearly sort out the fibrotic mice.

Paper Details

Date Published: 7 February 2007
PDF: 10 pages
Proc. SPIE 6470, Organic Photonic Materials and Devices IX, 647005 (7 February 2007); doi: 10.1117/12.713576
Show Author Affiliations
M. Strupler, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
A.-M. Pena, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
M. Hernest, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
Ctr. de Rechereche Cardiovasculaire Inserm-Lariboisière, INSERM (France)
P.-L. Tharaux, Ctr. de Rechereche Cardiovasculaire Inserm-Lariboisière, INSERM (France)
A. Fabre, INSERM, Univ. Paris 7 (France)
J. Marchal-Somme, INSERM, Univ. Paris 7 (France)
B. Crestani, INSERM, Univ. Paris 7 (France)
D. Débarre, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
J.-L. Martin, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
E. Beaurepaire, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)
M.-C. Schanne-Klein, Lab. for Optics and Biosciences, CNRS, Ecole Polytechnique (France)


Published in SPIE Proceedings Vol. 6470:
Organic Photonic Materials and Devices IX
James G. Grote; Francois Kajzar; Nakjoong Kim, Editor(s)

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