Approaches to altering particle distributions in cryo-electron microscopy sample preparation
Year: 2018
Type: article
Abstract: Cryo-electron microscopy (cryo-EM) can now be used to determine high-resolution structural information on a diverse range of biological specimens. Recent advances have been driven primarily by developments in microscopes and detectors, and through advances in image-processing software. However, for many single-particle cryo-EM projects, major bottlenecks currently remain at the sample-preparation stage; obtaining cryo-EM grids of sufficient quality for high-resolution single-particle analysis can require the careful optimization of many variables. Common hurdles to overcome include problems associated with the sample itself (buffer components, labile complexes), sample distribution (obtaining the correct concentration, affinity for the support film), preferred orientation, and poor reproducibility of the grid-making process within and between batches. This review outlines a number of methodologies used within the electron-microscopy community to address these challenges, providing a range of approaches which may aid in obtaining optimal grids for high-resolution data collection.
Language: English
Subfield: Structural Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $1,416
Awards: New infrastructure for EM studies at the University of Leeds., G3351971231, Structural and functional studies in indanomycin biosynthesis, Untangling the processes of replication and encapsidation in Picornavirales, Modern Cryo-Electron Microscopy with Direct Electron Detection at the University of Leeds., New infrastructure for EM studies at the University of Leeds., High-resolution structural studies of BK polyomavirus by cryo-electron microscopy, Modern Cryo-Electron Microscopy with Direct Electron Detection at the University of Leeds., Developing Electron Microscopy as a Tool in Structure-Based Drug Design
OpenAlex ID: W2804636455