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A simple pressure-assisted method for MicroED specimen preparation

Jingjing Zhao, Hongyi Xu, Hugo Lebrette, Marta Carroni, Helena Taberman, Martin Högbom, Xiaodong Zou
doi: https://doi.org/10.1101/665448
Jingjing Zhao
1Department of Materials and Environmental Chemistry, Stockholm University, Stockholm, Sweden
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Hongyi Xu
1Department of Materials and Environmental Chemistry, Stockholm University, Stockholm, Sweden
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  • For correspondence: hongyi.xu@mmk.su.se
Hugo Lebrette
3Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden
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Marta Carroni
2Science for Life Laboratory, Stockholm University, Solna, Sweden
3Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden
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Helena Taberman
4Max-Delbrück-Centrum for Molecular Medicine, Berlin, Germany
5Macromolecular Crystallography, Helmholtz-Zentrum Berlin für Materialien und Energie, Berlin, Germany
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Martin Högbom
3Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden
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Xiaodong Zou
1Department of Materials and Environmental Chemistry, Stockholm University, Stockholm, Sweden
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Abstract

Micro-crystal electron diffraction (MicroED) has shown great potential for structure determination of macromolecular crystals too small for X-ray diffraction. However, specimen preparation remains a major bottleneck. Here, we report a simple method for preparing MicroED specimens, named Preassis, in which excess liquid is removed through an EM grid with the assistance of pressure. We show the ice thicknesses can be controlled by tuning the pressure in combination with EM grids with appropriate hole sizes. Importantly, Preassis can handle a wide range of protein crystals grown in various buffer conditions including those with high viscosity, as well as samples with low crystal contents. Preassis is a simple and universal method for MicroED specimen preparation, and will significantly broaden the applications of MicroED.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵* e-mail: hongyi.xu{at}mmk.su.se, xzou{at}mmk.su.se

  • The updated manuscript focuses on MicroED specimen preparation only.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 19, 2021.
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A simple pressure-assisted method for MicroED specimen preparation
Jingjing Zhao, Hongyi Xu, Hugo Lebrette, Marta Carroni, Helena Taberman, Martin Högbom, Xiaodong Zou
bioRxiv 665448; doi: https://doi.org/10.1101/665448
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A simple pressure-assisted method for MicroED specimen preparation
Jingjing Zhao, Hongyi Xu, Hugo Lebrette, Marta Carroni, Helena Taberman, Martin Högbom, Xiaodong Zou
bioRxiv 665448; doi: https://doi.org/10.1101/665448

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