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Développement instrumental pour la microscopie de fluorescence résolue en temps: applications biomédicales

Abstract : For several biomedical issues, including neurobiology, imaging with high spatial and temporal resolution in a physiologically relevant cellular context is necessary. To address these requirements, we have developed an original setup which associates total internal reflection fluorescence (TIRF) and fluorescence lifetime imaging microscopy (FLIM). Indeed, FLIM allows a dynamic analysis of molecular and metabolic processes within cells. Combined with TIRF microscopy, which provides a sub-wavelength axial resolution, FLIM enables to analyze the activity of cell membrane, with efficient fluorescent background rejection.
The first aim of this project was to develop a fluorescence excitation source dedicated to lifetime measurement in time domain. We worked on a solution based on a solid state picosecond laser, and we used microstructured optical fibers to enlarge the spectrum by non linear effects. The second aspect of this work was the development and the characterization of the time-gated total internal reflection fluorescence microscope, which allows to measure fluorescence lifetime in wide field. Then, we used this setup to study using FRET probes the homodimerization of amyloid precursor protein (APP), which plays a crucial role in Alzheimer disease.
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https://pastel.archives-ouvertes.fr/tel-00376116
Contributor : Pierre Blandin <>
Submitted on : Thursday, April 16, 2009 - 4:40:52 PM
Last modification on : Friday, October 23, 2020 - 4:51:55 PM
Long-term archiving on: : Thursday, June 10, 2010 - 8:45:54 PM

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  • HAL Id : tel-00376116, version 1

Citation

Pierre Blandin. Développement instrumental pour la microscopie de fluorescence résolue en temps: applications biomédicales. Physique Atomique [physics.atom-ph]. Université Paris Sud - Paris XI, 2008. Français. ⟨tel-00376116⟩

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