title: Negative staining and cryo-negative staining of macromolecules and viruses for TEM.
abstract: In this review we cover the technical background to negative staining of biomolecules and viruses, and then expand upon the different possibilities and limitations. Topics range from conventional air-dry negative staining of samples adsorbed to carbon support films, the variant termed the "negative staining-carbon film" technique and negative staining of samples spread across the holes of holey-carbon support films, to a consideration of dynamic/time-dependent negative staining. For each of these approaches examples of attainable data are given. The cryo-negative staining technique for the specimen preparation of frozen-hydrated/vitrified samples is also presented. A detailed protocol to successfully achieve cryo-negative staining with ammonium molybdate is given, as well as examples of data, which support the claim that cryo-negative staining provides a useful approach for the high-resolution study of macromolecular and viral structure.
authors.affiliation: Department of Chemistry, and Institute for Macro Molecular Assemblies, The City College of CUNY, 160 Convent Ave, New York, NY, USA. sdecarlo@ccny.cuny.edu
authors.forename: Sacha
authors.initials: S
authors.lastname: De Carlo
authors.forename: J Robin
authors.initials: JR
authors.lastname: Harris
mesh_terms: D000388:Air; D005615:Freezing; D046911:Macromolecular Substances / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D016624:Negative Staining / Q000379:methods*; D014780:Viruses / Q000648:ultrastructure*
chemical_list: D046911:Macromolecular Substances