Negative staining and cryo-negative staining: applications in biology and medicine

Methods in Molecular Biology
J R Harris, Sacha De Carlo

Abstract

Negative staining is widely applicable to isolated viruses, protein molecules, macromolecular assemblies and fibrils, subcellular membrane fractions, liposomes and artificial membranes, synthetic DNA arrays, and also to polymer solutions and a variety of nanotechnology samples. Techniques are provided for the preparation of the necessary support films (continuous carbon and holey/perforated carbon). The range of suitable negative stains is presented, with some emphasis on the benefit of using ammonium molybdate and of negative stain-trehalose combinations. Protocols are provided for the single droplet negative staining technique (on continuous and holey carbon support films), the floating and carbon sandwich techniques in addition to the negative staining-carbon film (NS-CF) technique for randomly dispersed fragile molecules, 2D crystallization of proteins and for cleavage of cells and organelles. Immuno-negative staining and negative staining of affinity labeled complexes (e.g., biotin-streptavidin) are presented in some detail. The formation of immune complexes in solution for droplet negative staining is given, as is the use of carbon-plastic support films as an adsorption surface on which to perform immunolabeling or affini...Continue Reading

Citations

Feb 26, 2019·The Journal of Biological Chemistry·Dmitry Lyumkis
Mar 10, 2017·Scientific Reports·Reddy Ranjith K SamaDaryl A Bosco
Mar 4, 2021·Proceedings of the National Academy of Sciences of the United States of America·David S YangPeter G Vekilov
Apr 17, 2021·Scientific Reports·Alexandr B DuzhakYuri P Tsentalovich

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