Format

Send to

Choose Destination
J Struct Biol. 2013 Nov;184(2):193-202. doi: 10.1016/j.jsb.2013.09.003. Epub 2013 Sep 12.

Maximizing the potential of electron cryomicroscopy data collected using direct detectors.

Author information

1
Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037.
2
National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037.
3
Department of Computer Science and Engineering, University of California, San Diego, CA 92093.
#
Contributed equally

Abstract

Single-particle electron cryomicroscopy is undergoing a technical revolution due to the recent developments of direct detectors. These new recording devices detect electrons directly (i.e. without conversion into light) and feature significantly improved detective quantum efficiencies and readout rates as compared to photographic films or CCDs. We evaluated here the potential of one such detector (Gatan K2 Summit) to enable the achievement of near-atomic resolution reconstructions of biological specimens when coupled to a widely used, mid-range transmission electron microscope (FEI TF20 Twin). Compensating for beam-induced motion and stage drift provided a 4.4Å resolution map of Sulfolobus turreted icosahedral virus (STIV), which we used as a test particle in this study. Several motion correction and dose fractionation procedures were explored and we describe their influence on the resolution of the final reconstruction. We also compared the quality of this data to that collected with a FEI Titan Krios microscope equipped with a Falcon I direct detector, which provides a benchmark for data collected using a high-end electron microscope.

KEYWORDS:

Direct detectors; Electron microscopy; Near-atomic resolution; Sulfolobus turreted icosahedral virus

PMID:
24036281
PMCID:
PMC3832241
DOI:
10.1016/j.jsb.2013.09.003
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for PubMed Central
Loading ...
Support Center