[Microscopie de fluorescence de protéines autofluorescentes uniques pour la biologie cellulaire]
Nous présentons une revue sur l'utilisation des protéines autofluorescentes en microscopie de molécules uniques en biologie. Les caractéristiques photophysiques de plusieurs mutants de la protéine fluorescente verte (la « GFP ») ainsi que celles de la DsRed y sont comparées et discutées de manière critique en vue de leur utilisation dans des cellules vivantes. Des méthodes alternatives d'excitation, telle l'excitation biphotonique, ou d'analyses, telle la méthode de spectroscopie par corrélation de fluorescence sont envisagées. Nous rendons compte d'expériences utilisant la eGFP et préférentiellement la eYFP au niveau de la molécule unique dans des cellules vivantes. Nous reportons enfin, la première utilisation au niveau de la molécule individuelle de la citrine, un mutant de la eYFP plus résistant au photoblanchiment et son application à l'étude de la dynamique de récepteurs de neurotransmetteurs individuels dans la membrane de cellules vivantes.
In this paper we review the applicability of autofluorescent proteins for single-molecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and critically discussed for their use in cellular biology. The alternative use of two-photon excitation at the single-molecule level or Fluorescence Correlation Spectroscopy is envisaged for the study of individual autofluorescent proteins. Single-molecule experiments performed in live cells using eGFP and preferably eYFP fusion proteins are reviewed. Finally, the first use at the single-molecule level of citrine, a more photostable variant of the eYFP is reported when fused to a receptor for neurotransmitter in live cells.
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Mots-clés : détection de molécules uniques, GFP, DsRed, microscopie de fluorescence, excitation à deux photons, spectroscopie par corrélation de fluorescence
Laurent Cognet 1 ; Françoise Coussen 2 ; Daniel Choquet 2 ; Brahim Lounis 1
@article{CRPHYS_2002__3_5_645_0,
author = {Laurent Cognet and Fran\c{c}oise Coussen and Daniel Choquet and Brahim Lounis},
title = {Fluorescence microscopy of single autofluorescent proteins for cellular biology},
journal = {Comptes Rendus. Physique},
pages = {645--656},
publisher = {Elsevier},
volume = {3},
number = {5},
year = {2002},
doi = {10.1016/S1631-0705(02)01341-5},
language = {en},
}
TY - JOUR AU - Laurent Cognet AU - Françoise Coussen AU - Daniel Choquet AU - Brahim Lounis TI - Fluorescence microscopy of single autofluorescent proteins for cellular biology JO - Comptes Rendus. Physique PY - 2002 SP - 645 EP - 656 VL - 3 IS - 5 PB - Elsevier DO - 10.1016/S1631-0705(02)01341-5 LA - en ID - CRPHYS_2002__3_5_645_0 ER -
%0 Journal Article %A Laurent Cognet %A Françoise Coussen %A Daniel Choquet %A Brahim Lounis %T Fluorescence microscopy of single autofluorescent proteins for cellular biology %J Comptes Rendus. Physique %D 2002 %P 645-656 %V 3 %N 5 %I Elsevier %R 10.1016/S1631-0705(02)01341-5 %G en %F CRPHYS_2002__3_5_645_0
Laurent Cognet; Françoise Coussen; Daniel Choquet; Brahim Lounis. Fluorescence microscopy of single autofluorescent proteins for cellular biology. Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 645-656. doi : 10.1016/S1631-0705(02)01341-5. https://comptes-rendus.academie-sciences.fr/physique/articles/10.1016/S1631-0705(02)01341-5/
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