Immunohistochemical study of intermediate filament proteins on routinely processed, celloidin-embedded human temporal bone sections by using a new technique for antigen retrieval.
Although immunohistochemical studies of intermediate filament proteins have been carried out on temporal bone sections by using modified fixation/embedding techniques to preserve antigenicity, there have been no light microscopic studies concerning immunohistochemical staining on routinely formalin-fixed celloidin-embedded human temporal bone sections. A method for immunostaining routinely processed celloidin-embedded tissues would be extremely valuable in that it would permit study of the extensive collections of formalin-celloidin temporal bone specimens that exist in major centers of otopathologic research. Recently, we have developed a new technique which can be used to retrieve the antigenicity masked by formalin fixation and decalcification. This method requires immersing slides for 30 min at room temperature in a solution of saturated sodium hydroxide in methanol before immunostaining. Using this method, 45 celloidin-embedded human temporal bone sections were stained with monoclonal antibodies to keratin, vimentin, neurofilament, glial fibrillary acidic protein and desmin as primary antibodies using a sensitive streptavidin-biotin procedure. The results obtained by using this technique are at least equivalent to those obtained with modified fixatives, cryosections or immuno-electron microscopy. This new method may provide a useful approach for studying routinely processed, celloidin-embedded human temporal bone sections and open a new field in immuno-otopathology.
Shi
S R
SR
BioGenex Laboratories, San Ramon, CA 94583.
Tandon
A K
AK
Haussmann
R R
RR
Kalra
K L
KL
Taylor
C R
CR
D000328:Adult; D000911:Antibodies, Monoclonal; D003101:Collodion; D003893:Desmin / Q000032:analysis; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007633:Keratins / Q000032:analysis; D016900:Neurofilament Proteins / Q000032:analysis; D013701:Temporal Bone / Q000737:chemistry* / Q000473:pathology*; D016610:Tissue Embedding; D014746:Vimentin / Q000032:analysis
D000911:Antibodies, Monoclonal; D003893:Desmin; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; D014746:Vimentin; D007633:Keratins; D003101:Collodion