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Journal Article

rsEGFP2 enables fast RESOLFT nanoscopy of living cells.

MPS-Authors
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Grotjohann,  T.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Testa,  I.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Reuss,  M.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Brakemann,  T.
Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society;

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Eggeling,  C.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Jakobs,  S.
Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

1676138.pdf
(Publisher version), 4MB

Supplementary Material (public)
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Citation

Grotjohann, T., Testa, I., Reuss, M., Brakemann, T., Eggeling, C., Hell, S. W., et al. (2012). rsEGFP2 enables fast RESOLFT nanoscopy of living cells. eLife, 1: e00248. doi:10.7554/eLife.00248.


Cite as: http://hdl.handle.net/11858/00-001M-0000-000E-B7F9-E
Abstract
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.