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  Coordinate-targeted fluorescence nanoscopy with multiple off states.

Danzl, J. G., Sidenstein, S., Gregor, C., Urban, N. T., Ilgen, P., Jakobs, S., et al. (2016). Coordinate-targeted fluorescence nanoscopy with multiple off states. Nature Photonics, 10(2), 122-128. doi:10.1038/nphoton.2015.266.

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 Creators:
Danzl, J. G.1, Author           
Sidenstein, S.1, Author           
Gregor, C.1, Author           
Urban, N. T.1, Author           
Ilgen, P.1, Author           
Jakobs, S.2, Author           
Hell, S. W.1, 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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 Abstract: Far-field super-resolution fluorescence microscopy discerns fluorophores residing closer than the diffraction barrier by briefly transferring them in different (typically ON and OFF) states before detection. In coordinate-targeted super-resolution variants, such as stimulated emission depletion (STED) microscopy, this state difference is created by the intensity minima and maxima of an optical pattern, causing all fluorophores to assume the off state, for instance, except at the minima. Although strong spatial confinement of the on state enables high resolution, it also subjects the fluorophores to excess intensities and state cycles at the maxima. Here, we address these issues by driving the fluorophores into a second off state that is inert to the excess light. By using reversibly switchable fluorescent proteins as labels, our approach reduces bleaching and enhances resolution and contrast in live-cell STED microscopy. Using two or more transitions to off states is a useful strategy for augmenting the power of coordinate-targeted super-resolution microscopy.

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Language(s): eng - English
 Dates: 2016-01-182016-02
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1038/nphoton.2015.266
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Title: Nature Photonics
Source Genre: Journal
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Pages: - Volume / Issue: 10 (2) Sequence Number: - Start / End Page: 122 - 128 Identifier: -