| First Authors |
Andreas Müller
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| Authors |
Andreas Müller, Martin Neukam, Anna Ivanova, Anke Sönmez, Carla Münster, Susanne Kretschmar, Yannis Kalaidzidis, Thomas Kurth, Jean-Marc Verbavatz, Michele Solimena
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| Corresponding Authors |
Michele Solimena
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| Last Authors |
Michele Solimena
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| Journal Name |
Scientific reports (Sci Rep) |
| Volume |
7 |
| Issue |
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| Article Number |
23 |
| Open Access |
true |
| Print Publication Date |
2017-12-01 |
| Online Publication Date |
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| Abstract |
Correlative light and electron microscopy (CLEM) is a powerful approach to investigate the molecular ultrastructure of labeled cell compartments. However, quantitative CLEM studies are rare, mainly due to small sample sizes and the sensitivity of fluorescent proteins to strong fixatives and contrasting reagents for EM. Here, we show that fusion of a self-labeling protein to insulin allows for the quantification of age-distinct insulin granule pools in pancreatic beta cells by a combination of super resolution and transmission electron microscopy on Tokuyasu cryosections. In contrast to fluorescent proteins like GFP organic dyes covalently bound to self-labeling proteins retain their fluorescence also in epoxy resin following high pressure freezing and freeze substitution, or remarkably even after strong chemical fixation. This enables for the assessment of age-defined granule morphology and degradation. Finally, we demonstrate that this CLEM protocol is highly versatile, being suitable for single and dual fluorescent labeling and detection of different proteins with optimal ultrastructure preservation and contrast. |
| PDF |
Müller_2017_6796.pdf
(1.9 MB)
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| Cover Image |
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| Affiliated With |
EM Facility, Solimena, Zerial
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| Selected By |
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| Acknowledged Services |
Light Microscopy Facility
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| Publication Status |
Published |
| Edoc Link |
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| Sfx Link |
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| DOI |
10.1038/s41598-017-00033-x |
| PubMed ID |
28154417 |
| WebOfScience Link |
WOS:000396828900006 |
| Alternative Full Text URL |
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| Display Publisher Download Only |
false |
| Visible On MPI-CBG Website |
true |
| PDF Downloadable |
true |
| Created By |
thuem |
| Added Date |
2017-03-02 |
| Last Edited By |
brandl |
| Last Edited Date |
2017-06-08 12:10:25.389 |
| Library ID |
6796 |
| Document ID |
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| Entry Complete |
true |
| eDoc Compliant |
true |
| Include in Edoc Report |
true |
| In Pure |
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| Ready for eDoc Export |
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| Author Affiliations Complete |
false |
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