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

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:
 [1] ;  [1] ;  [1] ;  [1] ;  [2] ;  [3]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division
  2. 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
  3. 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:
Grant/Contract Number:
AC02-05CH11231; R01-GM083039; R01-GM065050
Type:
Accepted Manuscript
Journal Name:
Biophysical Journal
Additional Journal Information:
Journal Volume: 110; Journal Issue: 4; Journal ID: ISSN 0006-3495
Publisher:
Elsevier
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE; National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1379104