Reversibly switchable fluorescent proteins (RSFPs) are unusual and exciting proteins that may be reversibly switched by irradiation with light between a fluorescent (On) and a non-fluorescent (Off) state. They are enormously variable and therefore powerful probes for various forms of super-resolution microscopy including RESOLFT-microscopy. Using molecular design, we are changing and expanding the characteristics of RSFPs and investigating their properties on a molecular scale with a wide array of approaches including crystallography, Nuclear Magnetic Resonance (NMR), multi-dimension (MD) simulations and various forms of spectroscopy. We have created a palette of reversibly switching fluorescent proteins that cover a wide range of different colors, switching speeds and switching modes. These probes are used to study protein localization and dynamics on the nanoscale in living cells.
This endeavor is a joint effort with the Department of NanoBiophotonics at the Max Planck Institute for biophysical Chemistry in Göttingen.
Grotjohann, T.,Testa, I.,Leutenegger, M.,Bock, H.,Urban, N.T.,Lavoie-Cardinal, F.,Willig, K.I.,Eggeling, C.,Jakobs, S. and Hell, S.W.: "Diffraction-unlimited all-optical imaging and writing with a photochromic GFP"
Nature 478 (7368), 204-208. DOI 10.1038/nature10497Details
Brakemann, T.,Stiel, A.C.,Weber, G.,Andresen, M.,Testa, I.,Grotjohann, T.,Leutenegger, M.,Plessmann, U.,Urlaub, H.,Eggeling, C.,Wahl, M.C.,Hell, S.W. and Jakobs, S.: "A reversibly photoswitchable GFP-like protein with fluorescence excitation decoupled from switching"
Nature Biotech. 29 (10), 942-U132. DOI 10.1038/nbt.1952Details
Andresen, M.,Stiel, A.C.,Foelling, J.,Wenzel, D.,Schoenle, A.,Egner, A.,Eggeling, C.,Hell, S.W. and Jakobs, S.: "Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy"
Nature Biotech. 26 (9), 1035-1040. DOI 10.1038/nbt.1493Details