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TitleA simple pressure-assisted method for MicroED specimen preparation.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 5036, Year 2021
Publish dateAug 19, 2021
AuthorsJingjing Zhao / Hongyi Xu / Hugo Lebrette / Marta Carroni / Helena Taberman / Martin Högbom / Xiaodong Zou /
PubMed 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 ...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 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.
External linksNat Commun / PubMed:34413316 / PubMed Central
MethodsEM (single particle) / EM (tomography)
Resolution3.4 Å
Structure data

EMDB-10417:
GroEL map obtained using the Preassis method for grid preparation
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-10466:
Cryo-ET on GroEL
Method: EM (tomography)

Source
  • Escherichia coli (E. coli)
  • Escherichia coli K-12 (bacteria)

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