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Reversibly Photoswitchable Fluorescent Proteins for Live Cell Super-Resolution Microscopy

Vendredi 5 décembre 2014 11:00 - Duree : 1 heure
Lieu : Salle des séminaires de l’IBS - 71, avenue des Martyrs - Grenoble

Orateur : Stefan JAKOBS (Max Planck Institute for Biophysical Chemistry, Dept. of NanoBiophotonics & University Medical Center of Göttingen, Germany)

Reversibly switchable fluorescent proteins (RSFPs) have a similar overall molecular fold as the green fluorescent protein (GFP) and its derivatives. However, RSFPs can be repeatedly photoswitched between fluorescent and non-fluorescent states by irradiation with distinct light wavelengths. We generated a family of RSFPs exhibiting different switching modes and showing strong resistance against photobleaching and switching fatigue. Such properties make them particularly well suited for reversible saturable optical (fluorescence) transition (RESOLFT) super-resolution microscopy enabling the recording of raw data images from living cells and tissues with low levels of light. In conventional RSFPs, including Dronpa, rsEGFP and Padron, the molecular mechanism underlying the light driven switching is a cis-trans isomerization of the chromophore often accompanied by changes in the protonation state of the chromophore. As a consequence, the light wavelength used for fluorescence excitation also induces switching, which may represent a limitation for the usability of these RSFPs in some applications. With the RSFP Dreiklang we created an RSFP whose switching is disentangled from fluorescence readout. In Dreiklang, switching is based on a light induced reversible hydration/dehydration reaction that modifies the chromophore. Recent advances in the application and development of these probes as well as their use to overcome the diffraction barrier in RESOLFT and single molecule localization microscopy will be discussed.

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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