Factors that Influence the Formation and Stability of Thin,Cryo-EM Specimens |
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| Authors: | Robert?M. Glaeser Bong-Gyoon Han Roseann Csencsits Alison Killilea Arto Pulk Jamie?H.D. Cate |
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| Affiliation: | 1.Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California;2.Department of Molecular and Cell Biology and California Institute of Quantitative Biosciences, University of California, Berkeley, Berkeley, California;3.Department of Chemistry, University of California, Berkeley, Berkeley, California;4.Physical Biosciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, California |
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| 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. |
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