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Microwave-Accelerated Decalcification

Useful Methods for Research and Clinical Laboratories
  • Victoria J. Madden
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Morphological examination of calcified structures, by either light microscopy (LM) or electron microscopy (EM) often requires a decalcification (decal) step after fixation. Further processing steps must await the completion of the decal process. The time required for full decal with ethylenediamine tetraacetic acid (EDTA), a calcium (Ca) chelation agent, generally takes days to months, using routine methods. This time lag can be detrimental to both tissue morphology and antigenicity, and can hinder productivity and delay diagnostic results in clinical settings. Fortunately, decal times can be shortened dramatically with the use of a laboratory microwave (MW) oven (Keithley et al., 2000; Madden and Henson, 1997; Louw et al., 1994; Faria et al., 1992; Hellstrom and Nilsson, 1992; Ng and Ng, 1992; Roncaroli et al., 1991).

References

  1. Athanasou NA, Quinn J, Heryet A, Woods CG, McGee JO (1987) Effect of decalcification agents on immunoreactivity of cellular antigens. J Clin Pathol 40:874–878.PubMedCrossRefGoogle Scholar
  2. Baird IL, Winborn WB, Bockman DE (1967) Technique of decalcification suited to electron microscopy of tissues closely associated with bone. Anal Rec 159: 281–289.CrossRefGoogle Scholar
  3. Balaton AJ, Loget P (1989) Decalcification acceleree par les micro-ondes. Annal Pathol 9: 140–141.Google Scholar
  4. Dreiling FJ, Henson MM, Henson OW Jr (1999) Immunoelectron microscopic demonstration of collagen II in the basilar membrane and spiral ligament of New Zealand white rabbits (Abstract). Twenty Second Midwinter Res. Meeting, Assoc. Res. Otolaryngol. 22:77.Google Scholar
  5. Faria MR, Dechichi P, de Mello Rode S (1992) Decalcification of mineralized rat mandible tissues with the aid of microwaves. Rev Paul Med 110:283–284.PubMedGoogle Scholar
  6. Flower GC (1951) Notes on electrophoretic decalcification of bone. J Med Technol 9:106–107.Google Scholar
  7. Giberson RT, Demaree RS Jr. (1995) Microwave fixation: understanding the variables to achieve rapid and reproducible results. Microsc Res Tech 32:246–254.PubMedCrossRefGoogle Scholar
  8. Giberson RT, Demaree RS Jr., Nordhausen RW (1997) Four-hour processing of clinical/diagnostic specimens for electron microscopy using microwave technique. J Vet Diagn Invest 9:61–67.PubMedGoogle Scholar
  9. Hellström S, Nilsson M (1992) Microwave oven in temporal bone research. Acta Oto-Laryngol 493(Suppl): 15–18.Google Scholar
  10. Henson MM, Henson OW Jr. (2000a) Spiral limbus: specializations in Mormoopidae (Abstract) Twenty Third Midwinter Res. Meeting, Assoc. Res. Otolaryngol. 23:201.Google Scholar
  11. Henson OW Jr., Henson MM (2000b) Tympanic membrane: Highly developed smooth muscle arrays in the annulus fibrosus of mustached bats. J Assoc Res Otolaryngol 1:25–32.PubMedCrossRefGoogle Scholar
  12. Kaneko M, Tomita T, Nakase T, Takeuchi E, Iwasaki M, Sugamoto K, Yonenobu K, Ochi T (1999) Rapid decalcification using microwaves for in situ hybridization in skeletal tissues. Biotech Histochem 74:49–54.PubMedCrossRefGoogle Scholar
  13. Keithley EM, Truoung T, Chandronait B, Billings PB (2000) Immunohistochemistry and microwave decalcification of human temporal bones. Hearing Res 148:192–196.CrossRefGoogle Scholar
  14. Kok LP, Boon ME (1992) Microwave Cookbook for Microscopists. Coulomb, Leiden, The Netherlands.Google Scholar
  15. Login GR, Dvorak A (1994) The Microwave Toolbook: A Practical Guide for Microscopists. Beth Israel Hospital, Boston, MA.Google Scholar
  16. Louw I, De Beer DP, Du Plessis MJ (1994) Microwave histoprocessing of bone marrow trephine biopsies. Histochem J 26:487–494.PubMedCrossRefGoogle Scholar
  17. Madden VJ, Henson MM (1997) Rapid decalcification of temporal bones with preservation of ultrastructure. Hearing Res 111:76–84.CrossRefGoogle Scholar
  18. Morgan JM, Navabi H, Schmid KW, Jasani B (1994) Possible role of tissue-bound calcium ions in citrate-mediated high-temperature antigen retrieval. J Pathol 174:301–307.PubMedCrossRefGoogle Scholar
  19. Ng KH, Ng LL (1992) Microwave-stimulated decalcification of compact bones. EurJ Morphol 30:150–155.Google Scholar
  20. Roncaroli F, Mussa B, Bussolati G (1991) Microwave oven for improved tissue fixation and decalcification. Pathologica 83:307–310.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Victoria J. Madden
    • 1
  1. 1.Department of Pathology and Laboratory MedicineUniversity of North CarolinaChapel Hill

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