A simple pressure-assisted method for MicroED specimen preparation.

Zhao J, Xu H, Lebrette H, Carroni M, Taberman H, Högbom M, Zou X

Nat Commun 12 (1) 5036 [2021-08-19; online 2021-08-19]

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.

Cryo-EM [Collaborative]

PubMed 34413316

DOI 10.1038/s41467-021-25335-7

Crossref 10.1038/s41467-021-25335-7

pii: 10.1038/s41467-021-25335-7
pmc: PMC8377027

Publications 9.5.0