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  RESOLFT nanoscopy of fixed cells using a Z-domain based fusion protein for labelling.

Ilgen, P., Grotjohann, T., Jans, D. C., Kilisch, M., Hell, S. W., & Jakobs, S. (2015). RESOLFT nanoscopy of fixed cells using a Z-domain based fusion protein for labelling. PLoS One, 10(9): e0136233. doi:10.1371/journal.pone.0136233.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-4667-9 Version Permalink: http://hdl.handle.net/21.11116/0000-0002-AB34-3
Genre: Journal Article

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 Creators:
Ilgen, P.1, Author              
Grotjohann, T.2, Author              
Jans, D. C.1, Author              
Kilisch, M., Author
Hell, S. W.2, Author              
Jakobs, S.1, Author              
Affiliations:
1Research Group of Mitochondrial Structure and Dynamics, MPI for biophysical chemistry, Max Planck Society, ou_578566              
2Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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 Abstract: RESOLFT super-resolution microscopy allows subdiffraction resolution imaging of living cells using low intensities of light. It relies on the light-driven switching of reversible switchable fluorescent proteins (RSFPs). So far, RESOLFT imaging was restricted to living cells, because chemical fixation typically affects the switching characteristics of RSFPs. In this study we created a fusion construct (FLASR) consisting of the RSFP rsEGFP2 and the divalent form of the antibody binding Z domain from protein A. FLASR can be used analogous to secondary antibodies in conventional immunochemistry, facilitating simple and robust sample preparation. We demonstrate RESOLFT super-resolution microscopy on chemically fixed mammalian cells. The approach may be extended to other super-resolution approaches requiring fluorescent proteins in an aqueous environment.

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Language(s): eng - English
 Dates: 2015-09-16
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1371/journal.pone.0136233
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Title: PLoS One
Source Genre: Journal
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Pages: 11 Volume / Issue: 10 (9) Sequence Number: e0136233 Start / End Page: - Identifier: -