Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens

Glaeser, Robert M.; Han, Bong-Gyoon; Csencsits, Roseann; Killilea, Alison; Pulk, Arto; Cate, Jamie H. D.

doi:10.1016/j.bpj.2015.07.050

Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens

Journal Article · Thu Sep 17 04:00:00 EDT 2015 · Biophysical Journal

DOI:https://doi.org/10.1016/j.bpj.2015.07.050· OSTI ID:1379104

Glaeser, Robert M. ^[1]; Han, Bong-Gyoon ^[1]; Csencsits, Roseann ^[1]; Killilea, Alison ^[1]; Pulk, Arto ^[2]; Cate, Jamie H. D. ^[3]

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

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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1379104

Journal Information:: Biophysical Journal, Journal Name: Biophysical Journal Journal Issue: 4 Vol. 110; ISSN 0006-3495

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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