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J Mol Biol. 2016 Jan 29;428(2 Pt A):332-343. doi: 10.1016/j.jmb.2015.09.030. Epub 2015 Oct 9.

In Situ Cryo-Electron Tomography: A Post-Reductionist Approach to Structural Biology.

Author information

1
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
2
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany. Electronic address: baumeist@biochem.mpg.de.

Abstract

Cryo-electron tomography is a powerful technique that can faithfully image the native cellular environment at nanometer resolution. Unlike many other imaging approaches, cryo-electron tomography provides a label-free method of detecting biological structures, relying on the intrinsic contrast of frozen cellular material for direct identification of macromolecules. Recent advances in sample preparation, detector technology, and phase plate imaging have enabled the structural characterization of protein complexes within intact cells. Here, we review these technical developments and outline a detailed computational workflow for in situ structural analysis. Two recent studies are described to illustrate how this workflow can be adapted to examine both known and unknown cellular complexes. The stage is now set to realize the promise of visual proteomics--a complete structural description of the cell's native molecular landscape.

KEYWORDS:

cryo-EM; subtomogram averaging; template matching; tomography; visual proteomics

PMID:
26456135
DOI:
10.1016/j.jmb.2015.09.030
[Indexed for MEDLINE]

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