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research article

Diffraction-unlimited all-optical imaging and writing with a photochromic GFP

Grotjohann, Tim
•
Testa, Ilaria
•
Leutenegger, Marcel  
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2011
Nature

Lens-based optical microscopy failed to discern fluorescent features closer than 200 nm for decades, but the recent breaking of the diffraction resolution barrier by sequentially switching the fluorescence capability of adjacent features on and off is making nanoscale imaging routine. Reported fluorescence nanoscopy variants switch these features either with intense beams at defined positions or randomly, molecule by molecule. Here we demonstrate an optical nanoscopy that records raw data images from living cells and tissues with low levels of light. This advance has been facilitated by the generation of reversibly switchable enhanced green fluorescent protein (rsEGFP), a fluorescent protein that can be reversibly photoswitched more than a thousand times. Distributions of functional rsEGFP-fusion proteins in living bacteria and mammalian cells are imaged at <40-nanometre resolution. Dendritic spines in living brain slices are super-resolved with about a million times lower light intensities than before. The reversible switching also enables all-optical writing of features with subdiffraction size and spacings, which can be used for data storage.

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Type
research article
DOI
10.1038/nature10497
Author(s)
Grotjohann, Tim
Testa, Ilaria
Leutenegger, Marcel  
Bock, Hannes
Urban, Nicolai T.
Lavoie-Cardinal, Flavie
Willig, Katrin I.
Eggeling, Christian
Jakobs, Stefan
Hell, Stefan W.
Date Issued

2011

Published in
Nature
Volume

478

Start page

204

End page

208

Subjects

fluorescent protein

•

reversibly switchable

•

super-resolution microscopy

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
LOB  
Available on Infoscience
September 16, 2011
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/70967
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