Negative staining and cryo-negative staining of macromolecules and viruses for TEM.
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.
De Carlo
Sacha
S
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
Harris
J Robin
JR
D000388:Air; D005615:Freezing; D046911:Macromolecular Substances / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D016624:Negative Staining / Q000379:methods*; D014780:Viruses / Q000648:ultrastructure*
D046911:Macromolecular Substances