A simple pressure-assisted method for MicroED specimen preparation
| Autor: | Xiaodong Zou, Jingjing Zhao, Martin Högbom, Helena Taberman, Marta Carroni, Hugo Lebrette, Hongyi Xu |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Diffraction
Cancer Research Materials science Science Life Sciences Building Blocks of Life Structure and Function General Physics and Astronomy chemistry.chemical_element Biochemistry Article General Biochemistry Genetics and Molecular Biology Inorganic Chemistry Crystal Cryoelectron microscopy General Materials Science Physical and Theoretical Chemistry Composite material Multidisciplinary General Chemistry Condensed Matter Physics chemistry Electron diffraction Specimen preparation Structural biology Protein crystallization Carbon |
| Zdroj: | Nature Communications Nature Communications, Vol 12, Iss 1, Pp 1-7 (2021) |
| Popis: | 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. Micro-crystal electron diffraction (MicroED) has shown great potential for the structure determination of crystals that are too small for X-ray diffraction but MicroED sample preparation remains challenging. Here, the authors present Preassis, a pressure-assisted method for the preparation of MicroED specimens and demonstrate that Preassis can be applied to a wide range of protein crystal suspensions with low and high viscosities, as well as those with low crystal concentrations. |
| Databáze: | OpenAIRE |
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