Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins

Overview

Journal Sci Rep

Specialty Science

Date 2016 Jun 14

PMID 27292544

Citations 1

Authors

Chun-Ying Tsai

Yuan-Chih Chang

Ivan Lobato

Dirk Van Dyck

Fu-Rong Chen

Affiliations

Soon will be listed here.

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.

Citing Articles

Cryo-electron tomography related radiation-damage parameters for individual-molecule 3D structure determination.

Xue H, Zhang M, Liu J, Wang J, Ren G Front Chem. 2022; 10:889203.

PMID: 36110139 PMC: 9468540. DOI: 10.3389/fchem.2022.889203.

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