Microwave Processing Techniques for Electron Microscopy: A Four-Hour Protocol

  • Rick T. Giberson
  • Richard S. DemareeJr.
Part of the Methods in Molecular Biology™ book series (MIMB, volume 117)


The use of microwave energy to reduce processing times during fixation for electron microscopy (EM) is well established in the literature (1-5). The microwave protocol described in this chapter is based on the work of Giberson et al. (6,7) and differs from the microwave literature in that each step (fixation, dehydration, embedding, and resin polymerization) in specimen processing is done in the microwave. The time savings gained from this approach are considerable and the results, based on our experience and that of others (8,9), are equal or better than those achieved from routine processing protocols.


Microwave Irradiation Microcentrifuge Tube Propylene Oxide Sodium Cacodylate Cold Spot 
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  1. 1.
    Giberson, R. T. and Demaree, R. S., Jr. (1995) Microwave fixation: Understanding the variables to achieve rapid reproducible results. Microsc. Res. Tech. 32, 246–254.PubMedCrossRefGoogle Scholar
  2. 2.
    Benhamou, R., Noel, S., Grenier, J., and Asselin, A. (1991) Microwave energy fixation of plant tissue; an alternative approach that provides excellent preservation of ultrastructure and antigenicity. J. Electron Microsc. Tech. 17, 81–94.PubMedCrossRefGoogle Scholar
  3. 3.
    Login, G. R. and Dvorak, A. M. (1985) Microwave energy fixation for electron microscopy. Am. J. Pathol. 120, 230–243.PubMedGoogle Scholar
  4. 4.
    Login, G. R., Stavinoha, W. B., and Dvorak, A. M. (1986) Ultrafast microwave energy fixation for electron microscopy. J. Histochem. Cytochem. 34, 381–387.PubMedGoogle Scholar
  5. 5.
    Login, G. R., Dwyer, B. K., and Dvorak, A. M. (1990) Rapid primary microwave-osmium fixation. I. Preservation of structure for electron microscopy in seconds. J. Histochem. Cytochem. 38, 755–762.PubMedGoogle Scholar
  6. 6.
    Giberson, R. T., Demaree, R. S., Jr., and Nordhausen, R. W. (1997) Four-hour processing of clinical/diagnostic specimens for electron microscopy. J. Vet. Diagn. Invest. 9, 61–67.PubMedGoogle Scholar
  7. 7.
    Giberson, R. T., Smith, R. L., and Demaree, R. S. (1995) Three hour microwave tissue processing for transmission electron microscopy: From unfixed tissues to sections. Scanning 17(suppl. 5), 26–27.Google Scholar
  8. 8.
    Rassner, U. A., Crumrine, D. A., Nau, P., and Elias, P. M. (1997) Microwave incubation improves lipolytic enzyme preservation for ultrastructural cytochemistry. Histochem. J. 29, 387–392.PubMedCrossRefGoogle Scholar
  9. 9.
    Madden, V. J. and Henson, M. M. (1997) Rapid decalcification of temporal bones with preservation of ultrastructure. Hearing Research, 111, 76–84.PubMedCrossRefGoogle Scholar
  10. 10.
    Bozzola, J. J. and Russell, L. D. (1992) Electron Microscopy: Principles and Techniques for Biologists, Jones and Bartlett, Boston, MA, figure legends, p. 26.Google Scholar
  11. 11.
    Login, G. R., Galli, S. J., Morgan, E., Arizona, N., Schwartz, L. B., and Dvorak, A. M. (1987 a) Rapid microwave fixation of rat mast cells. I. Localization of granule chymase with an ultrastructural postembedding immunogold technique. J. Lab. Invest. 57(5), 592–599.Google Scholar
  12. 12.
    Login, G. R., Schnitt, S. J., and Dvorak, A. M. (1987 b) Methods in laboratory investigation: Rapid microwave fixation of human tissues for light microscopy immunoperoxidase identification of diagnostically useful antigens. J. Lab. Invest. 57(5), 585–591.Google Scholar
  13. 13.
    Login, G. R., Galli, S. J., and Dvorak, A. M. (1992) Immunocytochemical localization of histamine in secretory granules of rat peritoneal mast cells with conven-158 tional or rapid mcirowave fixation and an ultrastructural post-embedding immunogold technique. J. Histochem. Cytochem. 40(9), 1247–1256.PubMedGoogle Scholar
  14. 14.
    Leong, A. S.-Y., Milios, J., and Duncis, C. G. (1988) Antigen preservation in microwave-irradiated tissues: A comparison with formaldehyde fixation. J. Pathol. 156, 275–282.PubMedCrossRefGoogle Scholar
  15. 15.
    Cuevas, E. C., Bateman, A. C., Wilkins, B. S., Johnson, P. A., Williams, J. H., Lee, A. H. S., Jones, D. B., and Wright, D. H. (1994) Microwave antigen retrieval in immunocytochemistry: A study of 80 antibodies. J. Clin. Pathol. 47, 448–452.PubMedCrossRefGoogle Scholar
  16. 16.
    Shi, S., Key, M. E., and Kalra, K. L. (1991) Antigen retrieval in formalin-fixed, parafin-embedded tissues: An enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J. Histochem. Cytochem. 39(6), 741–748.PubMedGoogle Scholar
  17. 17.
    Estrada, J. C., Brinn, N. T., and Bossen, E. H. (1985) A rapid method of staining ultrathin sections for surgical pathology TEM with the use of the microwave oven. Brief Scientific Reports 83(5), 639–641.Google Scholar

Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Rick T. Giberson
    • 1
  • Richard S. DemareeJr.
    • 2
  1. 1.Ted Pella, IncRedding
  2. 2.Department of Biological SciencesCalifornia State UniversityChico

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