Imagerie Multimodale par Cartographie 3D en excitation pulsée : de la cellule au tissu

Abstract : To repair, supply or regenerate deficient organs, the bioengineering field consists of synthesising functionalised biomaterials. To visualise the synthesised structures in a non invasive way and in physiological conditions, new imaging techniques tend to be developed. Among them, microscopy associating multiphoton excitation with fluorescence detection or Second Harmonic Generation enables a visualisation in depth of extracellular matrix structures, without any exogenous dye. The first part of this work was to characterise multiphoton absorption probability in function of different excitation conditions, that means a modulation of the excitation beam through two different systems (Cavity Dumper and EOM). In a second part, we implemented and optimised the detection of the SHG signal coming from collagen in biological tissues. Through SHG measurements, we showed modifications occurring on the collagen network of the extracellular matrix of cartilage, when sample were submitted to mechanical (compression) or biochemical (enzymatic) constraint. We also were interested by the vascular research field and showed the ability of multiphoton microscopy to image without any fluorescent dye, the elastin network of the media and the collagen network of the adventice. We could appreciate qualitatively the effect of cryopreservation or fixation on the arterial wall, and the remodelling of a substitute implemented in a rabbit to supply its carotid. A last biological application concerned study of tumors, and showed us the complementarities between a macroscopic study with information obtained by microscopy.
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Elisabeth Werkmeister. Imagerie Multimodale par Cartographie 3D en excitation pulsée : de la cellule au tissu. Sciences agricoles. Université Henri Poincaré - Nancy 1, 2008. Français. ⟨NNT : 2008NAN10055⟩. ⟨tel-01748439⟩

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