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  Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection.

Winter, F., Loidolt, M., Westphal, V., Butkevich, A., Gregor, C., Sahl, S. J., et al. (2017). Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection. Scientific Reports, 7: 46492. doi:10.1038/srep46492.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-29CB-A Version Permalink: http://hdl.handle.net/21.11116/0000-0002-A70A-7
Genre: Journal Article

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 Creators:
Winter, F.1, Author              
Loidolt, M.1, Author              
Westphal, V.1, Author              
Butkevich, A.1, Author              
Gregor, C.1, Author              
Sahl, S. J.1, Author              
Hell, S. W.1, Author              
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1Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, ou_578627              

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 Abstract: The extension of fluorescence nanoscopy to larger numbers of molecular species concurrently visualized by distinct markers is of great importance for advanced biological applications. To date, up to four markers had been distinguished in STED experiments featuring comparatively elaborate imaging schemes and optical setups, and exploiting various properties of the fluorophores. Here we present a simple yet versatile STED design for multicolour imaging below the diffraction limit. A hyperspectral detection arrangement (hyperSTED) collects the fluorescence in four spectral channels, allowing the separation of four markers with only one excitation wavelength and a single STED beam. Unmixing of the different marker signals based on the simultaneous readout of all channels is performed with a non-negative matrix factorization algorithm. We illustrate the approach showing four-colour nanoscopy of fixed and living cellular samples.

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Language(s): eng - English
 Dates: 2017-04-18
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/srep46492
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Title: Scientific Reports
Source Genre: Journal
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Pages: 11 Volume / Issue: 7 Sequence Number: 46492 Start / End Page: - Identifier: -