Possibilities of Immunohistochemical Investigation on Human Temporal Bone

Abstract

After preliminary studies on human tonsillar tissue, lymph node tissue, or tissue from the appendix, it could be shown that after a fixation in Bouin solution or in sublimate-formaldehyde solution and subsequent embedding of the tissue in paraffinwax, the structure of the immunoglobulins present in the tissue remains intact. Even after long-term decalcifica-tion of the fixed tissue in EDTA, its structure does not alter and the immunoglobulins can be detected in high dilution either by the direct immunofluorescence method or with peroxidase-antiperoxidase. Human temporal bones from healthy individuals were fixed in Bouin solution or in sublimate-formaldehyde solution and incubated for sufficient time in EDTA decalcification solution until they were radiologically free of calcium. After embedding in paraffinwax or Paraplast, the immunoglobulins of the middle ear mucosa could be identified with the immunohistochemical methods described above. For the first time, localized immunoglobulins including the secretory component in the endolymphatic sac could also be demonstrated. The method is so sensitive that the binding of antibodies from the serum of patients with possibly immunopathological inner ear conditions can be detected by means of the indirect immunofluorescence test. Furthermore, other immunohistochemical assay methods can also be carried out with these methods (e.g. neuropeptide detection).

Key Words:

• decalcification
• preservation of antigenicity
• indirect immunofluorescence test