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  Coordinate-targeted and coordinate-stochastic super-resolution microscopy with the reversibly switchable fluorescent protein Dreiklang.

Jensen, N. A., Danzl, J. G., Willig, K. I., Lavoie-Cardinal, F., Brakemann, T., Hell, S. W., et al. (2014). Coordinate-targeted and coordinate-stochastic super-resolution microscopy with the reversibly switchable fluorescent protein Dreiklang. ChemPhysChem, 15(4), 756-762. doi:10.1002/cphc.201301034.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-82D9-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-E8D2-5
Genre: Journal Article

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1938101.pdf (Publisher version), 3MB
 
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 Creators:
Jensen, N. A.1, Author              
Danzl, J. G.1, Author              
Willig, K. I.1, Author              
Lavoie-Cardinal, F.1, Author              
Brakemann, T.2, Author              
Hell, S. W.1, Author              
Jakobs, S.2, Author              
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              
2Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              

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Free keywords: Dreiklang; fluorescent proteins; nanoscopy; photoswitching; resolution
 Abstract: Diffraction-unlimited far-field super-resolution fluorescence (nanoscopy) methods typically rely on transiently transferring fluorophores between two states, whereby this transfer is usually laid out as a switch. However, depending on whether this is induced in a spatially controlled manner using a pattern of light (coordinate-targeted) or stochastically on a single-molecule basis, specific requirements on the fluorophores are imposed. Therefore, the fluorophores are usually utilized just for one class of methods only. In this study we demonstrate that the reversibly switchable fluorescent protein Dreiklang enables live-cell recordings in both spatially controlled and stochastic modes. We show that the Dreiklang chromophore entails three different light-induced switching mechanisms, namely a reversible photochemical one, off-switching by stimulated emission, and a reversible transfer to a long-lived dark state from the S1 state, all of which can be utilized to overcome the diffraction barrier. We also find that for the single-molecule-based stochastic GSDIM approach (ground-state depletion followed by individual molecule return), Dreiklang provides a larger number of on-off localization events as compared to its progenitor Citrine. Altogether, Dreiklang is a versatile probe for essentially all popular forms of live-cell fluorescence nanoscopy.

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Language(s): eng - English
 Dates: 2014-02-042014-03-17
 Publication Status: Published in print
 Pages: 7
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1002/cphc.201301034
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Title: ChemPhysChem
Source Genre: Journal
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Pages: - Volume / Issue: 15 (4) Sequence Number: - Start / End Page: 756 - 762 Identifier: -