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Quispe J, Damiano J, Mick SE, et al. An improved holey carbon film for cryo-electron microscopy. Microsc Microanal. 2007;13(5):365-71doi: 10.1017/S1431927607070791.
Quispe, J., Damiano, J., Mick, S. E., Nackashi, D. P., Fellmann, D., Ajero, T. G., Carragher, B., & Potter, C. S. (2007). An improved holey carbon film for cryo-electron microscopy. Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada, 13(5), 365-71. https://doi.org/10.1017/S1431927607070791
Quispe, Joel, et al. "An improved holey carbon film for cryo-electron microscopy." Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada vol. 13,5 (2007): 365-71. doi: https://doi.org/10.1017/S1431927607070791
Quispe J, Damiano J, Mick SE, Nackashi DP, Fellmann D, Ajero TG, Carragher B, Potter CS. An improved holey carbon film for cryo-electron microscopy. Microsc Microanal. 2007 Oct;13(5):365-71. doi: 10.1017/S1431927607070791. PMID: 17900388.
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Microsc Microanal. 2007 Oct;13(5):365-71. doi: 10.1017/S1431927607070791.

An improved holey carbon film for cryo-electron microscopy.

Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada

Joel Quispe, John Damiano, Stephen E Mick, David P Nackashi, Denis Fellmann, Teddy G Ajero, Bridget Carragher, Clinton S Potter

Affiliations

  1. The National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, CA 92037, USA. [email protected]

PMID: 17900388 DOI: 10.1017/S1431927607070791

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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