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An improved holey carbon film for cryo-electron microscopy

Academic Article
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Overview

authors

  • Quispe, J.
  • Damiano, J.
  • Mick, S. E.
  • Nackashi, D. P.
  • Fellmann, D.
  • Ajero, T. G.
  • Carragher, Bridget
  • Potter, Clinton

publication date

  • October 2007

journal

  • Microscopy and Microanalysis  Journal

abstract

  • Two issues that often impact the cryo-electron microscopy (cryoEM) specimen preparation process are agglomeration of particles near hole edges in holey carbon films and variations in vitreous ice thickness. In many cases, the source of these issues was identified to be the residues and topography often seen in commercially available films. To study and minimize their impact during specimen preparation, an improved holey carbon film has been developed. Rather than using a consumable template based on soft materials that must be removed prior to grid assembly, a method was developed that uses a hard template and a water-soluble release layer to replicate the template pattern into the carbon films. The advantages of this method are the improved purity and flatness of the carbon films, and these attributes are shown to have a dramatic improvement on the distribution of single particles embedded in vitreous ice suspended across the holes. Improving particle distribution is an enabling factor toward increasing the throughput of data collection for cryoEM.
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Research

keywords

  • TEM
  • automation
  • cryo-electron microscopy
  • holey carbon films
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Identity

International Standard Serial Number (ISSN)

  • 1431-9276

Digital Object Identifier (DOI)

  • 10.1017/s1431927607070791

PubMed ID

  • 17900388
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Additional Document Info

start page

  • 365

end page

  • 371

volume

  • 13

issue

  • 5

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