skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

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

Journal Article · · Biophysical Journal
 [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
+ Show Author Affiliations

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.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE; National Institutes of Health (NIH)
Grant/Contract Number:
AC02-05CH11231; R01-GM083039; R01-GM065050
OSTI ID:
1379104
Journal Information:
Biophysical Journal, Vol. 110, Issue 4; ISSN 0006-3495
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 52 works
Citation information provided by
Web of Science

References (23)

Cryo-electron microscopy of viruses journal March 1984
Emerging techniques: Cryo-electron microscopy of vitrified biological specimens journal April 1985
Single-Particle Cryo-Electron Microscopy (Cryo-EM) book January 2014
2.2 A resolution cryo-EM structure of  -galactosidase in complex with a cell-permeant inhibitor journal May 2015
Alignment Error Envelopes for Single Particle Analysis journal February 2001
III. An essay on the cohesion of fluids journal January 1805
Phospholipid, Nature's own slide and cover slip for cryo-electron microscopy journal January 1989
Drying of solids wetted by thin liquid films journal September 1990
Possible mechanism for the spontaneous rupture of thin, free liquid films journal January 1966
How does a thin wetted film dry up? journal May 1994
Hydration forces: Observations, explanations, expectations, questions journal December 2011
Range and magnitude of the steric pressure between bilayers containing phospholipids with covalently attached poly(ethylene glycol) journal May 1995
Preparation of frozen-hydrated specimens for high resolution electron microscopy journal January 1984
Hydrophilic Support Films of Controlled Thickness and Composition journal October 1973
The fatty acid monolayer technique for preparing frozen-hydrated specimens journal January 1985
Molecular orientation of bacteriorhodopsin within the purple membrane of Halobacterium halobium. journal September 1978
TRPV1 structures in distinct conformations reveal activation mechanisms journal December 2013
Beam-induced motion correction for sub-megadalton cryo-EM particles journal August 2014
Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM journal May 2013
2.8 Å resolution reconstruction of the Thermoplasma acidophilum 20S proteasome using cryo-electron microscopy journal March 2015
Wetting and Capillary Phenomena of Water on Mica journal January 1998
Amphipols: Polymers that keep membrane proteins soluble in aqueous solutions journal December 1996
Structure of the TRPV1 ion channel determined by electron cryo-microscopy journal December 2013

Cited By (16)

Biological Applications at the Cutting Edge of Cryo-Electron Microscopy journal August 2018
Single particle cryo-EM reconstruction of 52 kDa streptavidin at 3.2 Angstrom resolution journal June 2019
Single particle electron cryomicroscopy: trends, issues and future perspective journal January 2016
Cryo-EM in drug discovery: achievements, limitations and prospects journal June 2018
A cryo-EM grid preparation device for time-resolved structural studies journal September 2019
Opinion: hazards faced by macromolecules when confined to thin aqueous films journal July 2016
Shake-it-off : A simple ultrasonic cryo-EM specimen preparation device journal May 2019
Manipulating and monitoring nanoparticles in micellar thin film superstructures journal December 2018
Routine Single Particle CryoEM Sample and Grid Characterization by Tomography journal December 2017
A Cryo-EM Grid Preparation Device for Time-Resolved Structural Studies journal March 2019
Routine single particle CryoEM sample and grid characterization by tomography text January 2018
Cryo-electron microscopy of an extremely halophilic microbe: technical aspects journal January 2017
Progress towards an optimal specimen support for electron cryomicroscopy journal April 2016
Size-Sorting and Pattern Formation of Nanoparticle-Loaded Micellar Superstructures in Biconcave Thin Films journal November 2017
Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation journal March 2020
Resilient quantum electron microscopy journal August 2022

Similar Records

Related Subjects

59 BASIC BIOLOGICAL SCIENCES