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

Ke Wang, Korrinn Strunk, Gongpu Zhao, Jennifer L. Gray, Peijun Zhang

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

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 2012

All Science Journal Classification (ASJC) codes

  • Structural Biology

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