Mario Perkovic, Michael Kunz, Ulrike Endesfelder, Stefanie Bunse, Christoph Wigge, Zhou Yu, Victor-Valentin Hodirnau, Margot Scheffer, Anja Seybert, Sebastian Malkusch, Erin Schuman, Mike Heilemann, Achilleas S. Frangakis
- Correlative microscopy incorporates the specificity of fluorescent protein labeling into high-resolution electron micrographs. Several approaches exist for correlative microscopy, most of which have used the green fluorescent protein (GFP) as the label for light microscopy. Here we use chemical tagging and synthetic fluorophores instead, in order to achieve protein-specific labeling, and to perform multicolor imaging. We show that synthetic fluorophores preserve their post-embedding fluorescence in the presence of uranyl acetate. Post-embedding fluorescence is of such quality that the specimen can be prepared with identical protocols for scanning electron microscopy (SEM) and transmission electron microscopy (TEM); this is particularly valuable when singular or otherwise difficult samples are examined. We show that synthetic fluorophores give bright, well-resolved signals in super-resolution light microscopy, enabling us to superimpose light microscopic images with a precision of up to 25 nm in the x-y plane on electron micrographs. To exemplify the preservation quality of our new method we visualize the molecular arrangement of cadherins in adherens junctions of mouse epithelial cells.
MetadatenAuthor: | Mario Perkovic, Michael Kunz, Ulrike EndesfelderORCiDGND, Stefanie Bunse, Christoph Wigge, Zhou Yu, Victor-Valentin HodirnauORCiD, Margot SchefferORCiDGND, Anja SeybertGND, Sebastian MalkuschORCiDGND, Erin SchumanORCiDGND, Mike HeilemannORCiDGND, Achilleas S. FrangakisORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-348115 |
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DOI: | https://doi.org/10.1016/j.jsb.2014.03.018 |
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ISSN: | 1095-8657 |
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ISSN: | 1047-8477 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/24698954 |
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Parent Title (English): | Journal of structural biology |
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Publisher: | Elsevier |
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Place of publication: | San Diego, Calif. |
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Document Type: | Article |
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Language: | English |
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Year of Completion: | 2014 |
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Date of first Publication: | 2014/03/31 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2014/10/27 |
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Tag: | Adherens junctions; Correlative electron and light microscopy; Electron tomography |
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Volume: | 186 |
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Issue: | 2 |
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Page Number: | 9 |
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First Page: | 205 |
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Last Page: | 213 |
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Note: | This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
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HeBIS-PPN: | 366621548 |
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Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
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| Angeschlossene und kooperierende Institutionen / MPI für Hirnforschung |
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Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
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Sammlungen: | Universitätspublikationen |
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Licence (German): | Creative Commons - Namensnennung-Nicht kommerziell-Keine Bearbeitung 3.0 |
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