Factors affecting macromolecule orientations in thin films formed in cryo-EM.

ABSTRACT: The formation of a vitrified thin film embedded with randomly oriented macromolecules is an essential prerequisite for cryogenic sample electron microscopy. Most commonly, this is achieved using the plunge-freeze method first described nearly 40 years ago. Although this is a robust method, the behaviour of different macromolecules shows great variation upon freezing and often needs to be optimized to obtain an isotropic, high-resolution reconstruction. For a macromolecule in such a film, the probability of encountering the air-water interface in the time between blotting and freezing and adopting preferred orientations is very high. 3D reconstruction using preferentially oriented particles often leads to anisotropic and uninterpretable maps. Currently, there are no general solutions to thi

SUBMITTER: Yadav S 

PROVIDER: S-EPMC11220838 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature