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Methods. 2016 May 1;100:3-15. doi: 10.1016/j.ymeth.2016.02.017. Epub 2016 Feb 28.

An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology.

Author information

1
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom. Electronic address: R.F.Thompson@leeds.ac.uk.
2
MLW Consulting, 11 Race Hill, Launceston, Cornwall PL15 9BB, United Kingdom.
3
Electron Bio-Imaging Centre, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, United Kingdom.
4
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.
5
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom. Electronic address: N.A.Ranson@leeds.ac.uk.

Abstract

Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a 'resolution revolution', owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM.

KEYWORDS:

Electron microscopy

PMID:
26931652
PMCID:
PMC4854231
DOI:
10.1016/j.ymeth.2016.02.017
[Indexed for MEDLINE]
Free PMC Article

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