3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography

Ke Wang, Korrinn Strunk, Gongpu Zhao, Jennifer Lynn Gray, Peijun Zhang

Research output: Contribution to journalArticle

38 Citations (Scopus)

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"

Original languageEnglish (US)
Pages (from-to)318-326
Number of pages9
JournalJournal of Structural Biology
Volume180
Issue number2
DOIs
StatePublished - Nov 1 2012

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Electron Microscope Tomography
Microtomy
Prokaryotic Cells
Eukaryotic Cells
Cellular Structures
Cell Communication
Artifacts
Ions
Viruses
Temperature

All Science Journal Classification (ASJC) codes

  • Structural Biology

Cite this

Wang, Ke ; Strunk, Korrinn ; Zhao, Gongpu ; Gray, Jennifer Lynn ; Zhang, Peijun. / 3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography. In: Journal of Structural Biology. 2012 ; Vol. 180, No. 2. pp. 318-326.
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3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography. / Wang, Ke; Strunk, Korrinn; Zhao, Gongpu; Gray, Jennifer Lynn; Zhang, Peijun.

In: Journal of Structural Biology, Vol. 180, No. 2, 01.11.2012, p. 318-326.

Research output: Contribution to journalArticle

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