title: Single-particle cryo-electron microscopy of macromolecular complexes.
abstract: Recent technological breakthroughs in image acquisition have enabled single-particle cryo-electron microscopy (cryo-EM) to achieve near-atomic resolution structural information for biological complexes. The improvements in image quality coupled with powerful computational methods for sorting distinct particle populations now also allow the determination of compositional and conformational ensembles, thereby providing key insights into macromolecular function. However, the inherent instability and dynamic nature of biological assemblies remain a tremendous challenge that often requires tailored approaches for successful implementation of the methodology. Here, we briefly describe the fundamentals of single-particle cryo-EM with an emphasis on covering the breadth of techniques and approaches, including low- and high-resolution methods, aiming to illustrate specific steps that are crucial for obtaining structural information by this method.
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, University of Michigan, 210 Washtenaw Avenue, Ann Arbor, MI 48109, USA skinioti@umich.edu dsouth@umich.edu.
authors.forename: Georgios
authors.initials: G
authors.lastname: Skiniotis
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, University of Michigan, 210 Washtenaw Avenue, Ann Arbor, MI 48109, USA skinioti@umich.edu dsouth@umich.edu.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Southworth
mesh_terms: D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods; D046911:Macromolecular Substances / Q000737:chemistry*; D008958:Models, Molecular; D008968:Molecular Conformation; D013048:Specimen Handling / Q000379:methods*
chemical_list: D046911:Macromolecular Substances
keywords: cryo-EM; macromolecular structure; negative-stain EM; single-particle EM