Publication Type |
Journal Article |
School or College |
College of Science |
Department |
Biology |
Creator |
Jorgensen, Erik |
Other Author |
Watanabe, Shigeki; Punge, Annedore; Hollopeter, Gunther; Willig, Katrin I.; Hobson, Robert John; Davis, M. Wayne; Hell, Stefan W. |
Title |
Protein localization in electron micrographs using fluorescence nanoscopy |
Date |
2010-01-01 |
Description |
A complete portrait of a cell requires a detailed description of its molecular topography: proteins must be linked to particular organelles. Immuno-electron microscopy can reveal locations of proteins with nanometer resolution but is limited by the quality of fixation, the paucity of antibodies, and the inaccessibility of the antigens. Here, we describe a method for correlative fluorescence electron microscopy (fEM) for the localization of proteins in electron micrographs. Proteins tagged with Citrine or tdEos were expressed in Caenorhabditis elegans, fixed and embedded. Tagged proteins were imaged from ultrathin sections using stimulated emission depletion microscopy (STED) or photoactivated localization microscopy (PALM). Fluorescence was correlated with organelles imaged in electron micrographs from the same sections. These methods were used to successfully localize histones, a mitochondrial protein, and a presynaptic dense projection protein in electron micrographs. |
Type |
Text |
Publisher |
Nature Publishing Group |
DOI |
doi:10.1038/nmeth.1537 |
Dissertation Institution |
University of Utah |
Language |
eng |
Bibliographic Citation |
Watanabe, S., Punge, A., Hollopeter, G., Willig, K. I., Hobson, R. J., Davis, M. W., Hell, S. W., & Jorgensen, E. M. (2010). Protein localization in electron micrographs using fluorescence nanoscopy. Nature Methods. |
Rights Management |
(c)Nature Publishing Group; Reprinted from Watanabe, S., Punge, A., Hollopeter, G., Willig, K. I., Hobson, R. J., Davis, M. W., Hell, S. W., & Jorgensen, E. M. (2010). Protein localization in electron micrographs using fluorescence nanoscopy. Nature Methods. |
Format Medium |
application/pdf |
Format Extent |
3,463,267 bytes |
Identifier |
uspace,14945 |
ARK |
ark:/87278/s6514gxt |
Setname |
ir_uspace |
ID |
707570 |
Reference URL |
https://collections.lib.utah.edu/ark:/87278/s6514gxt |