Pena, Ana-Maria (2006) Collagen second harmonic generation and application to the study of fibrosis using multiphoton microscopy. PhD thesis LOB, EP - LOB Laboratoire d'Optique et Biosciences, EP/X p.148.

Alternative Locations: http://www.imprimerie.polytechnique.fr/Theses/Files/Pena.pdf

Abstract

This work aimed at studying the second h!
armonic generation from collagen, the main constituent of the extracellular matrix of a wide variety of tissues and organs, and its application to the study of fibrosis using multiphoton microscopy, an optical imaging technique which has the unique ability of providing three-dimensional micrometer resolution within thick tissues. We first performed polarisation-resolved surface second harmonic generation experiments on thin films of collagen I and IV molecules, which allowed us to demonstrate that the SHG microscopy is a sensitive probe of collagen macromolecular organization and not of the collagen type. We next applied these results to the study of pulmonary and renal fibrosis in animal models, a pathological accumulation of fibrillar collagen which can progress to a terminal renal insufficiency or to a respiratory insufficiency often lethal eventually. We demonstrated that multiphoton microscopy permit to visualise the morphology of these tissues, to evidence all the characteri!
stic features of these pathologies and quantitatively evaluate the remodelling of the extracellular matrix during fibrosis. Finally, we have proposed fibrosis scores based on volume fractions of significant SHG signals which allowed us to appreciate the role of different factors (cells/assembling enzymes) in fibrosis. This methodology will help to develop new therapeutic approaches based on a better understanding of these fibrotic pathologies.

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