rsEGFP2 enables fast RESOLFT nanoscopy of living cells
Tim Grotjohann,
Ilaria Testa,
Matthias Reuss,
Tanja Brakemann,
Christian Eggeling,
Stefan W Hell,
Stefan Jakobs
Affiliations
Tim Grotjohann
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Ilaria Testa
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Matthias Reuss
Abberior Instruments GmbH, Göttingen, Germany; Science for Life Laboratory, Advanced Light Microscopy, Karolinska Institutet, Solna, Sweden
Tanja Brakemann
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Christian Eggeling
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Stefan W Hell
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Stefan Jakobs
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany; Department of Neurology, University of Göttingen, Göttingen, Germany
The super-resolution microscopy called RESOLFT relying on fluorophore switching between longlived states, stands out by its coordinate-targeted sequential sample interrogation using low light levels. While RESOLFT has been shown to discern nanostructures in living cells, the reversibly photoswitchable green fluorescent protein (rsEGFP) employed in these experiments was switched rather slowly and recording lasted tens of minutes. We now report on the generation of rsEGFP2 providing faster switching and the use of this protein to demonstrate 25–250 times faster recordings.
