Microscopie non linéaire de tissus biologiques : excitation multicouleur, faisceaux de Bessel, et excitation en nappe de lumière

Abstract : This work aimed at developing novel strategies in nonlinear microscopy to increase the number of avalaible contrasts and the imaging speed. Firstly, we investigated the use of synchronized pulse trains for multimodal nonlinear microscopy. We showed that this configuration allows the simultaneous and effective two photon excitation of blue, green-yellow and red fluorescent proteins. We have applied this technique to image large volume of Brainbow labeled tissues to develop strategies for mapping neural networks and reconstructing cell lineage trees in the central nervous system. More generally, we showed that synchronized pulse trains provide access to a broad range of nonlinear optical effects such as: two photon excited fluorescence (2PEF), second (SHG) and third harmonic (THG) generation and four-wave mixing (FWM). We then investigated strategies to increase the imaging frame rate of nonlinear microscopes. For this purpose, we first designed and built optical schemes that rely on Bessel beams in order to increase the depth of field of raster scanning microscopes. Finally, to potentially increase the imaging frame rate without degrading the optical sectionning we developped a nonlinear light sheet microscope with programmable excitation beam shape. Using this microscope, we compared the imaging properties of different excitation profiles (Gauss and Bessel) for applications to embryo imaging.
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https://pastel.archives-ouvertes.fr/pastel-00840186
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Submitted on : Monday, July 1, 2013 - 5:56:41 PM
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Pierre Mahou. Microscopie non linéaire de tissus biologiques : excitation multicouleur, faisceaux de Bessel, et excitation en nappe de lumière. Optique [physics.optics]. Ecole Polytechnique X, 2012. Français. ⟨NNT : 2597046289B⟩. ⟨pastel-00840186⟩

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