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Nat Methods. 2017 Aug;14(8):793-796. doi: 10.1038/nmeth.4347. Epub 2017 Jul 3.

Addressing preferred specimen orientation in single-particle cryo-EM through tilting.

Author information

1
The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, New York, USA.
2
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA.
3
Department of Integrative Structural and Computational Biology, Department of Chemistry, and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
4
Laboratory of Genetics and Helmsley Center for Genomic Medicine, The Salk Institute for Biological Studies, La Jolla, California, USA.

Abstract

We present a strategy for tackling preferred specimen orientation in single-particle cryogenic electron microscopy by employing tilts during data collection. We also describe a tool to quantify the resulting directional resolution using 3D Fourier shell correlation volumes. We applied these methods to determine the structures at near-atomic resolution of the influenza hemagglutinin trimer, which adopts a highly preferred specimen orientation, and of ribosomal biogenesis intermediates, which adopt moderately preferred orientations.

PMID:
28671674
PMCID:
PMC5533649
DOI:
10.1038/nmeth.4347
[Indexed for MEDLINE]
Free PMC Article

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