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

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. 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:

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

Publication Date:: Thu Sep 17 00:00:00 EDT 2015

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

Sponsoring Org.:: USDOE; National Institutes of Health (NIH)

OSTI Identifier:: 1379104

Grant/Contract Number:: AC02-05CH11231; R01-GM083039; R01-GM065050

Resource Type:: Accepted Manuscript

Journal Name:: Biophysical Journal

Additional Journal Information:: Journal Volume: 110; Journal Issue: 4; Journal ID: ISSN 0006-3495

Publisher:: Elsevier

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES

Citation Formats


                    Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, and Cate, Jamie H. D. Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens.  United States: N. p., 2015. 
Web.  doi:10.1016/j.bpj.2015.07.050.

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                    Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, & Cate, Jamie H. D. Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens.  United States.  https://doi.org/10.1016/j.bpj.2015.07.050

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                    Glaeser, Robert M., Han, Bong-Gyoon, Csencsits, Roseann, Killilea, Alison, Pulk, Arto, and Cate, Jamie H. D. Thu .  
"Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens".  United States.  https://doi.org/10.1016/j.bpj.2015.07.050.  https://www.osti.gov/servlets/purl/1379104.

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@article{osti_1379104,

  title        = {Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens},

  author       = {Glaeser, Robert M. and Han, Bong-Gyoon and Csencsits, Roseann and Killilea, Alison and Pulk, Arto and Cate, Jamie H. D.},

  abstractNote = {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.},

  doi          = {10.1016/j.bpj.2015.07.050},

  journal      = {Biophysical Journal},

  number       = 4,

  volume       = 110,

  place        = {United States},

  year         = {Thu Sep 17 00:00:00 EDT 2015},

  month        = {Thu Sep 17 00:00:00 EDT 2015}

}

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Works referencing / citing this record:

Biological Applications at the Cutting Edge of Cryo-Electron Microscopy
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Dillard, Rebecca S.; Hampton, Cheri M.; Strauss, Joshua D.
Microscopy and Microanalysis, Vol. 24, Issue 4
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Single particle cryo-EM reconstruction of 52 kDa streptavidin at 3.2 Angstrom resolution
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Fan, Xiao; Wang, Jia; Zhang, Xing
Nature Communications, Vol. 10, Issue 1
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Single particle electron cryomicroscopy: trends, issues and future perspective
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Vinothkumar, Kutti R.; Henderson, Richard
Quarterly Reviews of Biophysics, Vol. 49
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Cryo-EM in drug discovery: achievements, limitations and prospects
journal, June 2018

Renaud, Jean-Paul; Chari, Ashwin; Ciferri, Claudio
Nature Reviews Drug Discovery, Vol. 17, Issue 7
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A cryo-EM grid preparation device for time-resolved structural studies
journal, September 2019

Kontziampasis, Dimitrios; Klebl, David P.; Iadanza, Matthew G.
IUCrJ, Vol. 6, Issue 6
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Opinion: hazards faced by macromolecules when confined to thin aqueous films
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Glaeser, Robert M.; Han, Bong-Gyoon
Biophysics Reports, Vol. 3, Issue 1-3
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Shake-it-off : A simple ultrasonic cryo-EM specimen preparation device
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Rubinstein, John L.; Guo, Hui; Ripstein, Zev A.
Acta Crystallographica Section D Structural Biology
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Manipulating and monitoring nanoparticles in micellar thin film superstructures
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ten Hove, Jan Bart; van Leeuwen, Fijs W. B.; Velders, Aldrik H.
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Routine Single Particle CryoEM Sample and Grid Characterization by Tomography
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A Cryo-EM Grid Preparation Device for Time-Resolved Structural Studies
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Routine single particle CryoEM sample and grid characterization by tomography
text, January 2018

Noble, Alex J.; Dandey, Venkata P.; Wei, Hui
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Nature Communications, Vol. 9, Issue 1
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Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation
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Huang, Xiaojun; Zhang, Lei; Wen, Zuoling
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Resilient quantum electron microscopy
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Routine single particle CryoEM sample and grid characterization by tomography
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Noble, Alex J.; Dandey, Venkata P.; Wei, Hui
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Similar Records

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

Abstract

Citation Formats

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

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

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Routine Single Particle CryoEM Sample and Grid Characterization by Tomography journal, December 2017

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Cryo-electron microscopy of an extremely halophilic microbe: technical aspects journal, January 2017

Opinion: hazards faced by macromolecules when confined to thin aqueous films journal, July 2016

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

Manipulating and monitoring nanoparticles in micellar thin film superstructures journal, December 2018

Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation journal, March 2020

Resilient quantum electron microscopy journal, August 2022

Routine single particle CryoEM sample and grid characterization by tomography text, January 2018

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

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
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Opinion: hazards faced by macromolecules when confined to thin aqueous films
journal, July 2016

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
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Manipulating and monitoring nanoparticles in micellar thin film superstructures
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Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation
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Routine single particle CryoEM sample and grid characterization by tomography
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