RESOLFT Nanoscopy of Fixed Cells Using a Z-Domain Based Fusion Protein for Labelling
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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2267924"], "description"=>"<p>(A) Schematic of FLASR (ZZ-rsEGFP2<sub>tandem</sub>) bound to an immunoglobulin protein. (B-D) Maximum intensity projections of confocal z-stacks of methanol fixed mammalian CV-1 cells immunolabelled with antibodies against β-actin (B), vimentin (C) and α-tubulin (D). Subsequently, purified FLASR (red) was used to decorate the primary antibodies. Nuclei were stained with DAPI (blue). Scale bars: 50 μm.</p>", "links"=>[], "tags"=>["RSFP rsEGFP 2", "divalent form", "antibody binding Z domain", "FLASR", "protein", "sample preparation", "approach", "RESOLFT Nanoscopy", "microscopy", "fusion protein", "chemical fixation", "RESOLFT imaging", "subdiffraction resolution imaging"], "article_id"=>1546362, "categories"=>["Uncategorised"], "users"=>["Peter Ilgen", "Tim Grotjohann", "Daniel C. Jans", "Markus Kilisch", "Stefan W. Hell", "Stefan Jakobs"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0136233.g001", "stats"=>{"downloads"=>0, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunolabelling_with_FLASR_/1546362", "title"=>"Immunolabelling with FLASR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-16 03:36:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/2267948", "https://ndownloader.figshare.com/files/2267949"], "description"=>"<div><p>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.</p></div>", "links"=>[], "tags"=>["RSFP rsEGFP 2", "divalent form", "antibody binding Z domain", "FLASR", "protein", "sample preparation", "approach", "RESOLFT Nanoscopy", "microscopy", "fusion protein", "chemical fixation", "RESOLFT imaging", "subdiffraction resolution imaging"], "article_id"=>1546383, "categories"=>["Uncategorised"], "users"=>["Peter Ilgen", "Tim Grotjohann", "Daniel C. Jans", "Markus Kilisch", "Stefan W. Hell", "Stefan Jakobs"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0136233.s001", "https://dx.doi.org/10.1371/journal.pone.0136233.s002"], "stats"=>{"downloads"=>4, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_RESOLFT_Nanoscopy_of_Fixed_Cells_Using_a_Z_Domain_Based_Fusion_Protein_for_Labelling_/1546383", "title"=>"RESOLFT Nanoscopy of Fixed Cells Using a Z-Domain Based Fusion Protein for Labelling", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-09-16 03:36:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/2267945"], "description"=>"<p>Maximum intensity projections of confocal microscopy z-stacks of methanol fixed CV-1 cells immunolabelled with antibodies against β-actin (A) and vimentin (B). The purified recombinant fusion protein M-rsEGFP2<sub>tandem</sub> was used to decorate the primary antibodies (red). Nuclei were labelled with DAPI (blue). Scale bars: 50 μm.</p>", "links"=>[], "tags"=>["RSFP rsEGFP 2", "divalent form", "antibody binding Z domain", "FLASR", "protein", "sample preparation", "approach", "RESOLFT Nanoscopy", "microscopy", "fusion protein", "chemical fixation", "RESOLFT imaging", "subdiffraction resolution imaging"], "article_id"=>1546380, "categories"=>["Uncategorised"], "users"=>["Peter Ilgen", "Tim Grotjohann", "Daniel C. Jans", "Markus Kilisch", "Stefan W. Hell", "Stefan Jakobs"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0136233.g004", "stats"=>{"downloads"=>1, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunolabelling_with_an_M_rsEGFP2_tandem_fusion_protein_/1546380", "title"=>"Immunolabelling with an M-rsEGFP2<sub>tandem</sub> fusion protein.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-16 03:36:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/2267932"], "description"=>"<p>Comparison of RESOLFT super-resolution microscopy and the corresponding confocal microscopy images of CV-1 cells decorated with primary antibodies against vimentin (A), α-tubulin (B) and the nuclear pore complex protein Nup153 (C). (D) Line-profiles of the fluorescence intensities recorded between the arrowheads in (A-C), as indicated (confocal: light blue; RESOLFT: red). The line profiles in (1–3) are averaged across five adjacent line profiles that were perpendicular across the respective filament. The distance between two adjacent line profiles was the edge length of one pixel. Scale bars: 1 μm.</p>", "links"=>[], "tags"=>["RSFP rsEGFP 2", "divalent form", "antibody binding Z domain", "FLASR", "protein", "sample preparation", "approach", "RESOLFT Nanoscopy", "microscopy", "fusion protein", "chemical fixation", "RESOLFT imaging", "subdiffraction resolution imaging"], "article_id"=>1546370, "categories"=>["Uncategorised"], "users"=>["Peter Ilgen", "Tim Grotjohann", "Daniel C. Jans", "Markus Kilisch", "Stefan W. Hell", "Stefan Jakobs"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0136233.g002", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_RESOLFT_nanoscopy_of_methanol_fixed_cells_/1546370", "title"=>"RESOLFT nanoscopy of methanol fixed cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-16 03:36:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/2267937"], "description"=>"<p>The cell was decorated with primary antibodies against α-tubulin and FLASR. Scale bar: 5 μm.</p>", "links"=>[], "tags"=>["RSFP rsEGFP 2", "divalent form", "antibody binding Z domain", "FLASR", "protein", "sample preparation", "approach", "RESOLFT Nanoscopy", "microscopy", "fusion protein", "chemical fixation", "RESOLFT imaging", "subdiffraction resolution imaging"], "article_id"=>1546375, "categories"=>["Uncategorised"], "users"=>["Peter Ilgen", "Tim Grotjohann", "Daniel C. Jans", "Markus Kilisch", "Stefan W. Hell", "Stefan Jakobs"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0136233.g003", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_RESOLFT_super_resolution_image_of_an_entire_CV_1_cell_/1546375", "title"=>"RESOLFT super-resolution image of an entire CV-1 cell.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-16 03:36:56"}

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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