title: 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. Here we first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. 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. We thus suggest 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.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California. Electronic address: rmglaeser@lbl.gov.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Glaeser
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California.
authors.forename: Bong-Gyoon
authors.initials: BG
authors.lastname: Han
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California.
authors.forename: Roseann
authors.initials: R
authors.lastname: Csencsits
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California.
authors.forename: Alison
authors.initials: A
authors.lastname: Killilea
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California; Department of Molecular and Cell Biology and California Institute of Quantitative Biosciences, University of California, Berkeley, Berkeley, California.
authors.forename: Arto
authors.initials: A
authors.lastname: Pulk
authors.affiliation: Department of Molecular and Cell Biology and California Institute of Quantitative Biosciences, University of California, Berkeley, Berkeley, California; Department of Chemistry, University of California, Berkeley, Berkeley, California; Physical Biosciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California.
authors.forename: Jamie H D
authors.initials: JH
authors.lastname: Cate
mesh_terms: D000388:Air; D020285:Cryoelectron Microscopy; D004355:Drug Stability; D004583:Electrons; D011312:Pressure; D012997:Solvents / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry*; D014835:Volatilization; D014867:Water / Q000737:chemistry; D017155:Wettability
chemical_list: D012997:Solvents; D013501:Surface-Active Agents; D014867:Water