Fluorescent protein’s blinking : insights into a photophysical mechanism distorting super-resolution microscopy data
Vendredi 1er juillet 2016 11:00
- Duree : 1 heure
Lieu : Salle des séminaires de l’IBS - 71, avenue des Martyrs - Grenoble
Orateur : Romain BERARDOZZI (IBS/Groupe Dynamique et Cinétique des processus moléculaires)
Super-resolution microscopy based on single-molecule localization is a cutting edge technology allowing quantitative and dynamic studies of cellular events with nanometric accuracy. Nevertheless, the complex photophysical behavior of the fluorescent proteins used as markers in localization microscopy often leads to data distortion that may end up in misinterpretation of the biological process under study. In particular, multiple reversible transitions between fluorescent and non-fluorescent
states (“blinking”) may result in repeated counting of a single molecule, resulting in artifacts such as erroneous stoichiometries when characterizing biological assemblies. Based on a combination of single-molecule imaging, X-ray crystallography and optical spectroscopy, we set out to get better insights into the blinking mechanisms in photoconvertible fluorescent proteins such as mEos2 and Dendra2,
notably by elucidating the nature/structure of the non-fluorescent states. The results of our study reveal, surprisingly, that the blinking mechanisms in mEos2 and Dendra2 derivatives are mainly controlled
by the orientation of a single conserved amino acid (Arg66). These results open the door to the rational conception of optimized variants for quantitative and dynamics super-resolution microscopy.
Contact : ibs.seminaires@ibs.fr
Discipline évènement : (Biologie / Chimie)
Entité organisatrice : (IBS)
Nature évènement : (Séminaire)
Evènement répétitif : (Séminaire IBS)
Site de l'évènement : Polygone scientifique
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