Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules.

ABSTRACT: Cryo-electron microscopy (cryo-EM) is one of the primary methods used to determine the structures of macromolecules and their complexes. With the increased availability of cryo-electron microscopes, the preparation of high-quality samples has become a bottleneck in the cryo-EM structure-determination pipeline. Macromolecules can be damaged during the purification or preparation of vitrified samples for cryo-EM, making them prone to binding to the grid support, to aggregation or to the adoption of preferential orientations at the air-water interface. Here, it is shown that coating cryo-EM grids with a negatively charged polyelectrolyte, such as single-stranded DNA, before applying the sample reduces the aggregation of macromolecules and improves their distribution. The single-stranded DNA-c

SUBMITTER: Hrebik D 

PROVIDER: S-EPMC9629489 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature