Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy

Authors

  • Ana-Maria Pena,

    1. Laboratory for Optics and Biosciences, Ecole Polytechnique, 91128 Palaiseau, France
    2. CNRS, 91128 Palaiseau, France
    3. INSERM, U696, 91128 Palaiseau, France
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  • Aurélie Fabre,

    1. INSERM, U700, Paris, France
    2. Université Paris 7, Faculté de médecine X. Bichat, 75018 Paris, France
    3. AP-HP, Hôpital Bichat-Claude Bernard, services de pneumologie et laboratoire d'anatomie-pathologique, Paris, France
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  • Delphine Débarre,

    1. Laboratory for Optics and Biosciences, Ecole Polytechnique, 91128 Palaiseau, France
    2. CNRS, 91128 Palaiseau, France
    3. INSERM, U696, 91128 Palaiseau, France
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  • Joëlle Marchal-Somme,

    1. INSERM, U700, Paris, France
    2. Université Paris 7, Faculté de médecine X. Bichat, 75018 Paris, France
    3. AP-HP, Hôpital Bichat-Claude Bernard, services de pneumologie et laboratoire d'anatomie-pathologique, Paris, France
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  • Bruno Crestani,

    1. INSERM, U700, Paris, France
    2. Université Paris 7, Faculté de médecine X. Bichat, 75018 Paris, France
    3. AP-HP, Hôpital Bichat-Claude Bernard, services de pneumologie et laboratoire d'anatomie-pathologique, Paris, France
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  • Jean-Louis Martin,

    1. Laboratory for Optics and Biosciences, Ecole Polytechnique, 91128 Palaiseau, France
    2. CNRS, 91128 Palaiseau, France
    3. INSERM, U696, 91128 Palaiseau, France
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  • Emmanuel Beaurepaire,

    1. Laboratory for Optics and Biosciences, Ecole Polytechnique, 91128 Palaiseau, France
    2. CNRS, 91128 Palaiseau, France
    3. INSERM, U696, 91128 Palaiseau, France
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  • Marie-Claire Schanne-Klein

    Corresponding author
    1. Laboratory for Optics and Biosciences, Ecole Polytechnique, 91128 Palaiseau, France
    2. CNRS, 91128 Palaiseau, France
    3. INSERM, U696, 91128 Palaiseau, France
    • LOB, Ecole Polytechnique, Route de Saclay, 91128 Palaiseau cedex, France
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Abstract

The organization of collagen during fibrotic processes is poorly characterized because of the lack of appropriate methodologies. Here we show that multimodal multiphoton microscopy provides novel insights into lung fibrosis. We characterize normal and fibrotic pulmonary tissue in the bleomycin model, and show that second-harmonic generation by fibrillar collagen reveals the micrometer-scale three-dimensional spatial distribution of the fibrosis. We find that combined two-photon excited fluorescence and second-harmonic imaging of unstained lung tissue allows separating the inflammatory and fibrotic steps in this pathology, underlining characteristic features of fibroblastic foci in human Idiopathic Pulmonary Fibrosis samples. Finally, we propose phenomenological scores of lung fibrosis and we show that they unambiguously sort out control and treated mice, with a better sensitivity and reproducibility in the subpleural region. These results should be readily generalized to other organs, as an accurate method to assess extracellular matrix remodeling during fibrosis. Microsc. Res. Tech., 2007. © 2006 Wiley-Liss, Inc.

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