title: An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology.
abstract: Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a 'resolution revolution', owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM.
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom. Electronic address: R.F.Thompson@leeds.ac.uk.
authors.forename: Rebecca F
authors.initials: RF
authors.lastname: Thompson
authors.affiliation: MLW Consulting, 11 Race Hill, Launceston, Cornwall PL15 9BB, United Kingdom.
authors.forename: Matt
authors.initials: M
authors.lastname: Walker
authors.affiliation: Electron Bio-Imaging Centre, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, United Kingdom.
authors.forename: C Alistair
authors.initials: CA
authors.lastname: Siebert
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Muench
authors.affiliation: Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom. Electronic address: N.A.Ranson@leeds.ac.uk.
authors.forename: Neil A
authors.initials: NA
authors.lastname: Ranson
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020285:Cryoelectron Microscopy / Q000379:methods*; D005615:Freezing; D006801:Humans; D021621:Imaging, Three-Dimensional; D046911:Macromolecular Substances / Q000737:chemistry / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D058989:Vitrification
chemical_list: D046911:Macromolecular Substances
keywords: Electron microscopy