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  Live‐cell RESOLFT nanoscopy of transgenic Arabidopsis thaliana

Schnorrenberg, S., Ghareeb, H., Frahm, L., Grotjohann, T., Jensen, N., Teichmann, T., et al. (2020). Live‐cell RESOLFT nanoscopy of transgenic Arabidopsis thaliana. Plant direct, 4(9): e00261, pp. 1-8. doi:10.1002/pld3.261.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-F8B7-4 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-F8B8-3
Genre: Journal Article

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PlantDirect_4_2020_e00261.pdf (Any fulltext), 3MB
 
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 Creators:
Schnorrenberg, Sebastian, Author
Ghareeb, Hassan, Author
Frahm, Lars, Author
Grotjohann, Tim, Author
Jensen, Nickels, Author
Teichmann, Thomas, Author
Hell, Stefan W.1, Author              
Lipka, Volker, Author
Jakobs, Stefan, Author
Affiliations:
1Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society, ou_2364730              

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Free keywords: fluorescence lifetime gating, live cell imaging, plant, rsEGFP, super-resolution microscopy
 Abstract: Subdiffraction super‐resolution fluorescence microscopy, or nanoscopy, has seen remarkable developments in the last two decades. Yet, for the visualization of plant cells, nanoscopy is still rarely used. In this study, we established RESOLFT nanoscopy on living green plant tissue. Live‐cell RESOLFT nanoscopy requires and utilizes comparatively low light doses and intensities to overcome the diffraction barrier. We generated a transgenic Arabidopsis thaliana plant line expressing the reversibly switchable fluorescent protein rsEGFP2 fused to the mammalian microtubule‐associated protein 4 (MAP4) in order to ubiquitously label the microtubule cytoskeleton. We demonstrate the use of RESOLFT nanoscopy for extended time‐lapse imaging of cortical microtubules in Arabidopsis leaf discs. By combining our approach with fluorescence lifetime gating, we were able to acquire live‐cell RESOLFT images even close to chloroplasts, which exhibit very strong autofluorescence. The data demonstrate the feasibility of subdiffraction resolution imaging in transgenic plant material with minimal requirements for sample preparation.

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Language(s): eng - English
 Dates: 2020-07-202020-05-252020-08-032020-09-03
 Publication Status: Published online
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/pld3.261
 Degree: -

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Title: Plant direct
Source Genre: Journal
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Publ. Info: Hoboken, NJ : Wiley
Pages: - Volume / Issue: 4 (9) Sequence Number: e00261 Start / End Page: 1 - 8 Identifier: ISSN: 2475-4455