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Minimizing Crinkling of Soft Specimens Using Holey Gold Films on Molybdenum Grids for Cryogenic Electron Microscopy

Published online by Cambridge University Press:  04 June 2021

Xi Jiang Open the ORCID record for Xi Jiang [Opens in a new window] ,
Sunting Xuan ,
Ronald N. Zuckermann ,
Robert M. Glaeser ,
Kenneth H. Downing  and
Nitash P. Balsara
Xi Jiang*
Affiliation:
Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA
Sunting Xuan
Affiliation:
Molecular Foundry, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA
Ronald N. Zuckermann
Affiliation:
Molecular Foundry, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA
Robert M. Glaeser
Affiliation:
Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA
Kenneth H. Downing
Affiliation:
Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA
Nitash P. Balsara
Affiliation:
Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA94720, USA Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA94720, USA
*
*Author for correspondence: Xi Jiang, E-mail: xijiang@lbl.gov
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Abstract

We introduce a novel composite holey gold support that prevents cryo-crinkling and reduces beam-induced motion of soft specimens, building on the previously introduced all-gold support. The composite holey gold support for high-resolution cryogenic electron microscopy of soft crystalline membranes was fabricated in two steps. In the first step, a holey gold film was transferred on top of a molybdenum grid. In the second step, a continuous thin carbon film was transferred onto the holey gold film. This support (Au/Mo grid) was used to image crystalline synthetic polymer membranes. The low thermal expansion of Mo is not only expected to avoid cryo-crinkling of the membrane when the grids are cooled to cryogenic temperatures, but it may also act to reduce whatever crinkling existed even before cooling. The Au/Mo grid exhibits excellent performance with specimens tilted to 45°. This is demonstrated by quantifying beam-induced motion and differences in local defocus values. In addition, images of specimens on the Au/Mo grids that are tilted at 45° show high-resolution information of the crystalline membranes that, after lattice-unbending, extends beyond 1.5 Å in the direction perpendicular to the tilt axis.

Keywords

cryo-crinklingcryo-EMcrystalline polypeptoid membranescrystallographyspecimen flatness
Type
Software and Instrumentation
Information
Microscopy and Microanalysis , Volume 27 , Issue 4 , August 2021 , pp. 767 - 775
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Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press on behalf of the Microscopy Society of America

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Footnotes

†

Deceased.

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