Ultrastable gold substrates: Properties of a support for high-resolution electron cryomicroscopy of biological specimens

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• 作者：Christopher J. Russo ^{crusso@mrc-lmb.cam.ac.uk" class="auth_mail" title="E-mail the corresponding author} ; Lori A. Passmore ^{passmore@mrc-lmb.cam.ac.uk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Electron cryomicroscopy ; Single-particle reconstruction ; Electron tomography ; Gold supports ; Cryo-EM ; Protein structure
• 刊名：Journal of Structural Biology
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：193
• 期：1
• 页码：33-44
• 全文大小：3272 K

文摘

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.