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Spectroscopie de fluorescence dynamique confocale : réalisation du dispositif optique et application à l'étude de l'adsorption de protéines aux interfaces solide/liquide.

Abstract : The comprehension of the phenomena of adsorption of proteins to the solid-liquid interfaces is a key element in the design and the use of artificial membranes and biocompatible materials. In front of the complexity of the phenomena, the fundamental studies and their modelization have a strategic importance. The techniques of time resolved fluorescence and polarization of fluorescence permit et weak concentration to obtain information both the direct environment and the movement of a fluorescent molecule. In addition, the confocal fluorescence techniques make possible as for them the visualization of a volume of the order of the femtolitre and perfectly localised.
We developed an innovating optical device allowing to carry out studies of adsorption under flow by measurements of time resolved fluorescence and polarization of fluorescence in confocal mode.
The intrinsic fluorescence of proteins or direct labelin by fluorophore was not a simple system to study the movements of proteins. The solution was to carry out a marking of the avidine by using its strong affinity for the biotin. We synthesized a fluorescent probe composed of biotin ethylenediamine and alexa fluorine 594. This probe forms a very stable complex with the avidin.
Finally, we observed the evolution of the profile of concentration during the phenomenon of adsorption. We observed the kinetics of adsorption and the impact over the fluorescence lifetime and the correlation time of the avidine adsorbed on a model surface (mica) and on a hemodialysis membrane (AN-69).
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https://tel.archives-ouvertes.fr/tel-00011288
Contributor : Sébastien Balme <>
Submitted on : Tuesday, January 3, 2006 - 7:23:07 PM
Last modification on : Tuesday, May 28, 2019 - 2:18:02 PM
Long-term archiving on: : Tuesday, April 6, 2010 - 4:21:13 PM

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  • HAL Id : tel-00011288, version 1

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Sébastien Balme. Spectroscopie de fluorescence dynamique confocale : réalisation du dispositif optique et application à l'étude de l'adsorption de protéines aux interfaces solide/liquide.. Biophysique [physics.bio-ph]. Université Montpellier II - Sciences et Techniques du Languedoc, 2005. Français. ⟨tel-00011288⟩

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