Time resolved applications for Cryo-EM; approaches, challenges and future directions

David P Klebl; Louie Aspinall; Stephen P Muench

doi:10.1016/j.sbi.2023.102696

Time resolved applications for Cryo-EM; approaches, challenges and future directions

Curr Opin Struct Biol. 2023 Dec:83:102696. doi: 10.1016/j.sbi.2023.102696. Epub 2023 Sep 14.

Authors

David P Klebl¹, Louie Aspinall², Stephen P Muench³

Affiliations

¹ School of Biomedical Sciences, Astbury Centre for Structural Molecular Biology, University of Leeds, UK.
² School of Molecular and Cellular Biology, University of Leeds, UK.
³ School of Biomedical Sciences, Astbury Centre for Structural Molecular Biology, University of Leeds, UK. Electronic address: s.p.muench@leeds.ac.uk.

PMID: 37716094
DOI: 10.1016/j.sbi.2023.102696

Abstract

Developments within the cryo-EM field have allowed us to generate higher-resolution "static" structures and pull out different conformational states which exist at equilibrium within the sample. Moreover, to trap non-equilibrium states and determine conformations that are present after a defined period of time (typically in the ms time frame) new approaches have been developed for the application of time-resolved cryo-EM. Here we give an overview of these different approaches and the limitations and strengths of each whilst identifying some of the current challenges to achieve higher resolutions and trap states within faster time frames. Time-resolved applications may play an important role in the ever-expanding toolkit of cryo-EM and open up new possibilities in both single particle and tomographic studies.

Keywords: Cryo-EM; Protein dynamics; Sample preparation; Time-resolved.

Publication types

Review

MeSH terms

Cryoelectron Microscopy* / methods
Molecular Conformation