Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens
Abstract
Poor consistency of the ice thickness from one area of a cryo-electron microscope (cryo-EM) specimen grid to another, from one grid to the next, and from one type of specimen to another, motivates a reconsideration of how to best prepare suitably thin specimens. We first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. Furthermore, we then suggest that the importance of there being a surfactant monolayer at the air-water interface of thin, cryo-EM specimens has been largely underappreciated. In fact, a surfactant layer (of uncontrolled composition and surface pressure) can hardly be avoided during standard cryo-EM specimen preparation. Thus it is suggested that better control over the composition and properties of the surfactant layer may result in more reliable production of cryo-EM specimens with the desired thickness.
- Authors:
-
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division; Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. of Quantitative Biosciences
- Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. of Quantitative Biosciences, Dept. of Chemistry; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Physical Biosciences Division
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE; National Institutes of Health (NIH)
- OSTI Identifier:
- 1379104
- Grant/Contract Number:
- AC02-05CH11231; R01-GM083039; R01-GM065050
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biophysical Journal
- Additional Journal Information:
- Journal Volume: 110; Journal Issue: 4; Journal ID: ISSN 0006-3495
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, and Cate, Jamie H. D. Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens. United States: N. p., 2015.
Web. doi:10.1016/j.bpj.2015.07.050.
Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, & Cate, Jamie H. D. Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens. United States. https://doi.org/10.1016/j.bpj.2015.07.050
Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, and Cate, Jamie H. D. Thu .
"Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens". United States. https://doi.org/10.1016/j.bpj.2015.07.050. https://www.osti.gov/servlets/purl/1379104.
@article{osti_1379104,
title = {Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens},
author = {Glaeser, Robert M. and Han, Bong-Gyoon and Csencsits, Roseann and Killilea, Alison and Pulk, Arto and Cate, Jamie H. D.},
abstractNote = {Poor consistency of the ice thickness from one area of a cryo-electron microscope (cryo-EM) specimen grid to another, from one grid to the next, and from one type of specimen to another, motivates a reconsideration of how to best prepare suitably thin specimens. We first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. Furthermore, we then suggest that the importance of there being a surfactant monolayer at the air-water interface of thin, cryo-EM specimens has been largely underappreciated. In fact, a surfactant layer (of uncontrolled composition and surface pressure) can hardly be avoided during standard cryo-EM specimen preparation. Thus it is suggested that better control over the composition and properties of the surfactant layer may result in more reliable production of cryo-EM specimens with the desired thickness.},
doi = {10.1016/j.bpj.2015.07.050},
journal = {Biophysical Journal},
number = 4,
volume = 110,
place = {United States},
year = {Thu Sep 17 00:00:00 EDT 2015},
month = {Thu Sep 17 00:00:00 EDT 2015}
}
Web of Science
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