A simple pressure-assisted method for MicroED specimen preparation

dc.contributor.authorZhao, Jingjingen
dc.contributor.authorXu, Hongyien
dc.contributor.authorLebrette, Hugoen
dc.contributor.authorCarroni, Martaen
dc.contributor.authorTaberman, Helenaen
dc.contributor.authorHögbom, Martinen
dc.contributor.authorZou, Xiaodongen
dc.date.accessioned2025-06-11T09:34:26Z
dc.date.available2025-06-11T09:34:26Z
dc.date.issued2021-08-19en
dc.description.abstractMicro-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 carbon 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 concentrations. Preassis is a simple and universal method for MicroED specimen preparation, and will significantly broaden the applications of MicroED.en
dc.description.sponsorshipThe project is supported by the Knut and Alice Wallenberg Foundation (2012.0112 and 2018.0237, X.Z.; 2017.0275, M.H.), the Swedish Research Council (2017-04018, M.H.; 2017-05333, H.X.; 2019-00815, X.Z.) and the Science for Life Laboratory through the pilot project grant Electron Nanocrystallography, and MicroED@SciLifeLab. The authors acknowledge the Cryo-EM Swedish National Facility jointly funded by the Knut and Alice Wallenberg, Family Erling Persson and Kempe Foundations, SciLife-Lab, Stockholm University, and Umeå University. The authors also thank I. Schlichting at Max-Plank Institute for Medical Research for providing the tetragonal lysozyme sample.en
dc.description.statusPeer-revieweden
dc.format.extent7en
dc.identifier.issn2041-1723en
dc.identifier.otherPubMed:34413316en
dc.identifier.otherORCID:/0000-0002-8271-3906/work/184830370en
dc.identifier.scopus85113215299en
dc.identifier.urihttp://www.scopus.com/inward/record.url?scp=85113215299&partnerID=8YFLogxKen
dc.identifier.urihttps://hdl.handle.net/1885/733758434
dc.language.isoenen
dc.provenanceThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/ licenses/by/4.0/.en
dc.rights© 2021, The Author(s).en
dc.sourceNature Communicationsen
dc.titleA simple pressure-assisted method for MicroED specimen preparationen
dc.typeJournal articleen
dspace.entity.typePublicationen
local.contributor.affiliationZhao, Jingjing; Stockholm Universityen
local.contributor.affiliationXu, Hongyi; Stockholm Universityen
local.contributor.affiliationLebrette, Hugo; Stockholm Universityen
local.contributor.affiliationCarroni, Marta; Stockholm Universityen
local.contributor.affiliationTaberman, Helena; Max Delbrück Center for Molecular Medicine in the Helmholtz Associationen
local.contributor.affiliationHögbom, Martin; Stockholm Universityen
local.contributor.affiliationZou, Xiaodong; Stockholm Universityen
local.identifier.citationvolume12en
local.identifier.doi10.1038/s41467-021-25335-7en
local.identifier.pure5f3118a0-c142-4195-b8bb-ec9e009d0f30en
local.identifier.urlhttps://www.scopus.com/pages/publications/85113215299en
local.type.statusPublisheden

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