期刊
JOURNAL OF STRUCTURAL BIOLOGY
卷 193, 期 1, 页码 33-44出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2015.11.006
关键词
Electron cryomicroscopy; Single-particle reconstruction; Electron tomography; Gold supports; Cryo-EM; Protein structure
资金
- European Research Council under the European Union's Seventh Framework Programme (FP7)/ERC [261151]
- Medical Research Council [U105192715]
- Medical Research Council [MC_U105192715] Funding Source: researchfish
- MRC [MC_U105192715] Funding Source: UKRI
Electron cryomicroscopy (cryo-EM) allows structure determination of a wide range of biological molecules and specimens. All-gold supports improve cryo-EM images by reducing radiation-induced motion and image blurring. Here we compare the mechanical and electrical properties of all-gold supports to amorphous carbon foils. Gold supports are more conductive, and have suspended foils that are not compressed by differential contraction when cooled to liquid nitrogen temperatures. These measurements show how the choice of support material and geometry can reduce specimen movement by more than an order of magnitude during low-dose imaging. We provide methods for fabrication of all-gold supports and preparation of vitrified specimens. We also analyse illumination geometry for optimal collection of high resolution, low-dose data. Together, the support structures and methods herein can improve the resolution and quality of images from any electron cryomicroscope. (C) 2015 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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