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Negative Staining: a Valuable Technique for Studying Subcellular Components

Published online by Cambridge University Press:  02 July 2020

Sara E. Miller
Sara E. Miller*
Affiliation:
Departments of Microbiology and Pathology, Duke University Medical Center, Durham, NC, 27710
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Extract

Negative staining is the most frequently used procedure for preparing particulate specimens, e.g., cell organelles, macromolecules, and viruses, for electron microscopy (Figs. 1-4). The main advantage is that it is rapid, requiring only minutes of preparation time. Another is that it avoids some of the harsh chemicals, e.g., organic solvents, used in thin sectioning. Also, it does not require advanced technical skill. It is widely used in virology, both in classification of viruses as well as diagnosis of viral diseases. Notwithstanding the necessity for fairly high particle counts, virus identification by negative staining is advantageous in not requiring specific reagents such as antibodies, nucleic acid probes, or protein standards which necessitate prior knowledge of potential pathogens for selection of the proper reagent. Furthermore, it does not require viable virions as does growth in tissue culture. Another procedure that uses negative contrasting is ultrathin cryosectioning (Fig. 5).

In 1954 Farrant was the first to publish negatively stained material, ferritin particles.

Type
Technologists’ Forum: Special Topics and Symposium
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 341 - 342
Copyright
Copyright © Microscopy Society of America 1997

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References

1.Farrant, J.L., Biochim. Biophys. Acta 13(1954)569.10.1016/0006-3002(54)90376-5CrossRefGoogle Scholar
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6.Hayat, M.A. and Miller, S.E.,Negative Staining New York:McGraw-Hill Publishing Co. (1990).Google Scholar
7. I am grateful to Hopkins, Benfor computer and photography assistance and to Dr.Howell, David for critically reviewing this manuscript.Google Scholar