Schaffer, Miroslava and Engel, Benjamin D. and Laugks, Tim and Mahamid, Julia and Plitzko, Jürgen M. and Baumeister, Wolfgang. (2015) Cryo-focused ion beam sample preparation for imaging vitreous cells by cryo-electron tomography. Bio-protocol, 5 (17). e1575.
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Official URL: https://edoc.unibas.ch/89563/
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Abstract
Cryo-electron tomography (CET) is a well-established technique for imaging cellular and molecular structures at sub-nanometer resolution. As the method is limited to samples that are thinner than 500 nm, suitable sample preparation is required to attain CET data from larger cell volumes. Recently, cryo-focused ion beam (cryo-FIB) milling of plunge-frozen biological material has been shown to reproducibly yield large, homogeneously thin, distortion-free vitreous cross-sections for state-of-the-art CET. All eukaryotic and prokaryotic cells that can be plunge-frozen can be thinned with the cryo-FIB technique. Together with advances in low-dose microscopy, this has shifted the frontiers of in situ structural biology. In this protocol we describe the typical steps of the cryo-FIB technique, starting with fully grown cell cultures. Three recently investigated biological samples are given as examples.
Faculties and Departments: | 05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology and Biophysics (Engel) |
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UniBasel Contributors: | Engel, Ben |
Item Type: | Article, refereed |
Article Subtype: | Research Article |
Publisher: | Bio-Protocol |
e-ISSN: | 2331-8325 |
Note: | Publication type according to Uni Basel Research Database: Journal article |
Language: | English |
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Last Modified: | 23 Aug 2022 07:49 |
Deposited On: | 23 Aug 2022 07:49 |
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