Fluorescence microscopy of single autofluorescent proteins for cellular biology

Laurent Cognet (1) , Françoise Coussen (2) , Daniel Choquet (2) , Brahim Lounis (1)

1 CPMOH - Centre de physique moléculaire optique et hertzienne
2 PCS - Physiologie cellulaire de la synapse

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.

Mots clés: single-molecule; GFP; DsRed; fluorescence microscopy; two-photon excitation; fluorescence correlation spectroscopy.; fluorescence correlation spectroscopy

Domaines: Optique [physics.optics]; Biophysique [physics.bio-ph]

Laurent Cognet, Françoise Coussen, Daniel Choquet, Brahim Lounis. Fluorescence microscopy of single autofluorescent proteins for cellular biology. Comptes Rendus de l'Academie des Sciences. Série IV, Physique, Astronomie, 2002, 3, pp.645. ⟨hal-00143984⟩