Title |
Correlative in-resin super-resolution and electron microscopy using standard fluorescent proteins
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Published in |
Scientific Reports, March 2015
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DOI | 10.1038/srep09583 |
Pubmed ID | |
Authors |
Errin Johnson, Elena Seiradake, E. Yvonne Jones, Ilan Davis, Kay Grünewald, Rainer Kaufmann |
Abstract |
We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer superior structural preservation, performing in-resin super-resolution, however, remains a challenge. We identified key aspects of the sample preparation procedure of high pressure freezing, freeze substitution and resin embedding that are critical for preserving fluorescence and photo-switching of standard fluorescent proteins, such as mGFP, mVenus and mRuby2. This enabled us to combine single molecule localization microscopy with transmission electron microscopy imaging of standard fluorescent proteins in cryo-fixed resin embedded cells. We achieved a structural resolution of 40-50 nm (~17 nm average single molecule localization accuracy) in the fluorescence images without the use of chemical fixation or special fluorophores. Using this approach enabled the correlation of fluorescently labeled structures to the ultrastructure in the same cell at the nanometer level and superior structural preservation. |
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United Kingdom | 3 | 43% |
United States | 1 | 14% |
France | 1 | 14% |
Unknown | 2 | 29% |
Demographic breakdown
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Scientists | 2 | 29% |
Mendeley readers
Geographical breakdown
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Portugal | 2 | <1% |
Czechia | 2 | <1% |
Germany | 1 | <1% |
Netherlands | 1 | <1% |
France | 1 | <1% |
Sweden | 1 | <1% |
Switzerland | 1 | <1% |
Finland | 1 | <1% |
United Kingdom | 1 | <1% |
Other | 2 | <1% |
Unknown | 218 | 94% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 52 | 23% |
Student > Master | 18 | 8% |
Student > Bachelor | 16 | 7% |
Professor | 10 | 4% |
Other | 35 | 15% |
Unknown | 38 | 16% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 52 | 23% |
Engineering | 15 | 6% |
Physics and Astronomy | 9 | 4% |
Chemistry | 9 | 4% |
Other | 37 | 16% |
Unknown | 38 | 16% |