Kaplan, M., Nicolas, W. J., Xhao, W., Carter, S. D. , Metskas, L. A., Chreifi, G., Ghosal, D. and Jensen, G. J. (2020) In situ imaging and structure determination of biomolecular complexes using electron cryo-tomography. In: Gonen, T. and Nannenga, B. L. (eds.) CryoEM: Methods and Protocols. Series: Methods in molecular biology (2215). Humana: New York, NY, pp. 83-111. ISBN 9781071609651 (doi: 10.1007/978-1-0716-0966-8_4)
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Abstract
Electron cryo-tomography (cryo-ET) is a technique that allows the investigation of intact macromolecular complexes while they are in their cellular milieu. Over the years, cryo-ET has had a huge impact on our understanding of how large biomolecular complexes look like, how they assemble, disassemble, function, and evolve(d). Recent hardware and software developments and combining cryo-ET with other techniques, e.g., focused ion beam milling (FIB-milling) and cryo-light microscopy, has extended the realm of cryo-ET to include transient molecular complexes embedded deep in thick samples (like eukaryotic cells) and enhanced the resolution of structures obtained by cryo-ET. In this chapter, we will present an outline of how to perform cryo-ET studies on a wide variety of biological samples including prokaryotic and eukaryotic cells and biological plant tissues. This outline will include sample preparation, data collection, and data processing as well as hybrid approaches like FIB-milling, cryosectioning, and cryo-correlated light and electron microscopy (cryo-CLEM).
| Item Type: | Book Sections |
|---|---|
| Status: | Published |
| Glasgow Author(s) Enlighten ID: | Carter, Dr Stephen |
| Authors: | Kaplan, M., Nicolas, W. J., Xhao, W., Carter, S. D., Metskas, L. A., Chreifi, G., Ghosal, D., and Jensen, G. J. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research |
| Publisher: | Humana |
| ISBN: | 9781071609651 |
| Published Online: | 27 December 2020 |
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