Title
Hollow cone electron imaging for single particle 3D reconstruction of proteins Hollow cone electron imaging for single particle 3D reconstruction of proteins
Author
Faculty/Department
Faculty of Sciences. Physics
Publication type
article
Publication
London :Nature Publishing Group ,
Subject
Physics
Source (journal)
Scientific reports. - London, 2011, currens
Volume/pages
6(2016) , p. 1-9
ISSN
2045-2322
Article Reference
27701
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
The main bottlenecks for high-resolution biological imaging in electron microscopy are radiation sensitivity and low contrast. The phase contrast at low spatial frequencies can be enhanced by using a large defocus but this strongly reduces the resolution. Recently, phase plates have been developed to enhance the contrast at small defocus but electrical charging remains a problem. Single particle cryo-electron microscopy is mostly used to minimize the radiation damage and to enhance the resolution of the 3D reconstructions but it requires averaging images of a massive number of individual particles. Here we present a new route to achieve the same goals by hollow cone dark field imaging using thermal diffuse scattered electrons giving about a 4 times contrast increase as compared to bright field imaging. We demonstrate the 3D reconstruction of a stained GroEL particle can yield about 13.5 Å resolution but using a strongly reduced number of images.
Full text (open access)
https://repository.uantwerpen.be/docman/irua/2e44ef/134038.pdf
https://repository.uantwerpen.be/docman/irua/7cd31b/134038.pdf
Handle