Hollow cone electron imaging for single particle 3D reconstruction of proteins

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review

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Author(s)

  • Chun-Ying Tsai
  • Yuan-Chih Chang
  • Ivan Lobato
  • Dirk Van Dyck
  • Fu-Rong Chen

Detail(s)

Original languageEnglish
Article number27701
Journal / PublicationScientific Reports
Volume6
Early online date13 Jun 2016
Publication statusPublished - 2016
Externally publishedYes

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.

Citation Format(s)

Hollow cone electron imaging for single particle 3D reconstruction of proteins. / Tsai, Chun-Ying; Chang, Yuan-Chih; Lobato, Ivan; Van Dyck, Dirk; Chen, Fu-Rong.

In: Scientific Reports, Vol. 6, 27701, 2016.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review