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Microscopie non linéaire : chiralité et génération de second harmonique

Abstract : Nonlinear microscopy : chirality and second harmonic generation Nonlinear microscopy enables functional 3D imaging of living thick tissues or cells. The contrast in imaging may come from synthetic dyes (exogenous contrast) or from proteins in tissues (endogenous contrast). We perform nonlinear microscopy with both of these approaches. In particularly, we use SHG (Second Harmonic Generation) and TPEF (Two Photon Excited Fluorescence). Our work on dyes for SHG microscopy rests on nonlinear optical study of the mechanims of optical activity in chiral molecules. The chirality of dyes should enhance SHG membrane’s imagery when molecules are in centrosymetric arrangement in membranes. We demonstrate that chirality of the dye must come from an excitonic coupling. We study such a colorant : the ASTB (Acridin Substituted Troger Base). We study and use two sources of endogenous contrast : collagen (SHG) and elastin (TPEF). After characterisation of their nonlinear optical properties, we demonstrate that the combination of TPEF and SHG in microscopy enables a morphological study of fresh living arterial tissue in Rats. We also show the dramatic effects of lindane (an insecticide) on the vascular system. We perform the first nanotoxinologic study in nonlinear microscopy based on SHG signal from myosin filaments. We measure a contracture of atrial frog cells with the introduction of a toxin (saxitoxin) with a resolution of 20 nm.
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  • HAL Id : tel-01234992, version 1



Thierry Boulesteix. Microscopie non linéaire : chiralité et génération de second harmonique. Optique [physics.optics]. Paris-Sud XI, 2004. Français. ⟨NNT : 2004PA112052⟩. ⟨tel-01234992⟩



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