DOI: 10.1017/s1431927607070791. An Improved Holey Carbon Film for Cryo-Electron Microscopy

An Improved Holey Carbon Film for Cryo-Electron Microscopy

10.1017/s1431927607070791

Crossref journal-article

Oxford University Press (OUP)

Microscopy and Microanalysis (286)

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.

Bibliography

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, 13(5), 365â371.

Authors 8

Joel Quispe (first)
John Damiano (additional)
Stephen E. Mick (additional)
David P. Nackashi (additional)
Denis Fellmann (additional)
Teddy G. Ajero (additional)
Bridget Carragher (additional)
Clinton S. Potter (additional)

References 9 Referenced 54

Dates

Type	When
Created	17 years, 10 months ago (Sept. 28, 2007, 6:39 a.m.)
Deposited	2 years, 6 months ago (Feb. 14, 2023, 3:39 p.m.)
Indexed	3 months, 1 week ago (May 12, 2025, 10:25 a.m.)
Issued	17 years, 10 months ago (Sept. 28, 2007)
Published	17 years, 10 months ago (Sept. 28, 2007)
Published Online	17 years, 10 months ago (Sept. 28, 2007)
Published Print	17 years, 10 months ago (Oct. 1, 2007)

Funders 0

None

BibTeX

@article{Quispe_2007, title={An Improved Holey Carbon Film for Cryo-Electron Microscopy}, volume={13}, ISSN={1435-8115}, url={http://dx.doi.org/10.1017/s1431927607070791}, DOI={10.1017/s1431927607070791}, number={5}, journal={Microscopy and Microanalysis}, publisher={Oxford University Press (OUP)}, author={Quispe, Joel and Damiano, John and Mick, Stephen E. and Nackashi, David P. and Fellmann, Denis and Ajero, Teddy G. and Carragher, Bridget and Potter, Clinton S.}, year={2007}, month=sep, pages={365–371} }

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  "abstract": "<jats:p>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.</jats:p>",
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