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Microwave Processing of Cell Monolayers in Situ for Postembedding Immunocytochemistry with Retention of Ultrastructure and Antigenicity

Published online by Cambridge University Press:  02 July 2020

Victoria J. Madden
Victoria J. Madden*
Affiliation:
Department of Pathology and Laboratory Medicine, The University of North Carolina at Chapel Hill, CB #7525 Brinkhous-Bullitt Bldg., Chapel Hill, N.C.27599-7525
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Extract

Increasingly, cells isolated from blood and body fluids and cells grown in culture are becoming the experimental models of choice for biological research. The demand for demonstrating biochemical processes morphologically is also becoming commonplace in the electron microscopy laboratory. Successful fixation, in situ embedment, and ultrathinsectioning of cell monolayers can be difficult to achieve for routine transmission electron microscopy. For postembedding immunocytochemistry, processing becomes more complex due to fixation constraints and the use of acrylic resins. The object of this paper is to present a reliable, rapid method for processing monolayers that preserves both the ultrastructure of the cells and antigenicity.

The equipment used for this procedure was a Pelco Model 3440 MAX laboratory microwave oven equipped with a temperature probe and a maximum power output of 800 watts. Using a neon bulb array, the oven cavity is calibrated to determine the microwave energy distribution.

Type
Specimen Preparation
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 854 - 855
Copyright
Copyright © Microscopy Society of America

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References

1.Login, G.R. and Dvorak, A.. The Microwave Toolbook: A Practical Guide for Microscopists, Beth Israel Hospital, Boston, MA (1994) 51–61.Google Scholar
2.Giberson, R.T. and Demaree, R.S., Jr. Microwave fixation: understanding the variables to achieve rapid and reproducible results, Microsc. Res. Tech. 32 (1995) 246–254.CrossRefGoogle Scholar
3. This research was supported by the UNC Department of Pathology and Laboratory Medicine. Thank you to Dr. C.R. Bagnell, Jr. for mentoring, and to Dr. Katherine Pryzwansky and Dr. Nadia Malouf for supplying cells and antibodies.Google Scholar