NDLI: 3D Structure Determination of Native Mammalian Cells using Cryo-FIB and Cryo-electron Tomography

Content Provider	PubMed Central
Author	Wang, Ke Strunk, Korrinn Zhao, Gongpu Gray, Jennifer L. Zhang, Peijun
Copyright Year	2012
Abstract	Cryo-electron tomography (cryo-ET) has enabled high resolution three-dimensional (3D) structural analysis of virus and host cell interactions and many cell signaling events; these studies, however, have largely been limited to very thin, peripheral regions of eukaryotic cells or to small prokaryotic cells. Recent efforts to make thin, vitreous sections using cryo-ultramicrotomy have been successful, however, this method is technically very challenging and with many artifacts. Here, we report a simple and robust method for creating in situ, frozen-hydrated cell lamellas using a focused ion beam at cryogenic temperature (cryo-FIB), allowing access to any interior cellular regions of interest. We demonstrate the utility of cryo-FIB with high resolution 3D cellular structures from both bacterial cells and large mammalian cells. The method will not only facilitate high-throughput 3D structural analysis of biological specimens, but is also broadly applicable to sample preparation of thin films and surface materials without the need for FIB “lift-out”.
Related Links	http://dx.doi.org/10.1016/j.jsb.2012.07.003
Ending Page	326
Page Count	9
Starting Page	318
File Format	PDF
ISSN	10478477
e-ISSN	10958657
Journal	Journal of structural biology
Issue Number	2
Volume Number	180
Language	English
Publisher Date	2012-11-01
Access Restriction	Open
Subject Keyword	Structural Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Structural Biology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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