Negative staining and cryo-negative staining of macromolecules and viruses for TEM.

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

Micron (Oxford, England : 1993) 2011 Feb

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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. Copyright © 2009 Elsevier Ltd. All rights reserved.

Citation

Sacha De Carlo, J Robin Harris. Negative staining and cryo-negative staining of macromolecules and viruses for TEM. Micron (Oxford, England : 1993). 2011 Feb;42(2):117-31