Inner ear fine structures of the hamster in frozen sections used for immunohistochemical assays for inner ear diseases

Abstract

Frozen sections of the inner ear of the hamster enable detailed investigations of the fine structures in immunofluorescence assays. At high magnification single mitochondria can be identified by their reactions with an antiserum containing antibodies against mitochondria. In the positive reaction with an antiserum against nuclei, the typical green fluorescence is restricted to the nuclei, which are mostly separated by the surrounding cytoplasm. The method of immunohistochemical assay using frozen sections from the non-decalcified inner ear is very time-consuming and cannot be recommended for the routine diagnosis of inner ear diseases, although it may be useful in research and for studying critical clinical cases.

A new immunohistochemical method for the detection of gentamicin in inner ear fluid compartments

“…Temperature conditions were -25 ° C for the cryotome room and approximately -4 0°C for the cooling table [23]. After embedding in Tissue Tek, the cochlea was sectioned.…”

Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear

“…As a general rule, it could be considered that in unfixed and non-decalcified tissues all antigens are preserved structurally unmodified (Soliman 1988a,b;Bauwens et al 1990;Wei and Giebel 1991), at least for a brief period after obtaining the sample. Therefore, the use of non-decalcified cochleae was sometimes necessary to avoid deleterious effects of prolonged or aggressive decalcification processes (Soliman 1988a,b;Wei and Giebel 1991;Fujiyoshi et al 1992).…”

“…Therefore, the use of non-decalcified cochleae was sometimes necessary to avoid deleterious effects of prolonged or aggressive decalcification processes (Soliman 1988a,b;Wei and Giebel 1991;Fujiyoshi et al 1992). However, dissected or non-decalcified tissues did not allow for the preservation of the fine structural details of the auditory receptor, which usually appeared distorted (Soliman 1988a,b;Bauwens et al 1990;Tonnaer et al 1990;Wei and Giebel 1991;Fujiyoshi et al 1992). In frozen tissues (Wei and Giebel 1991), only some sections showed enough morphology to recognize tissue organization, but there was few cells even when liquid nitrogen was used (Soliman 1988a,b).…”

“…However, dissected or non-decalcified tissues did not allow for the preservation of the fine structural details of the auditory receptor, which usually appeared distorted (Soliman 1988a,b;Bauwens et al 1990;Tonnaer et al 1990;Wei and Giebel 1991;Fujiyoshi et al 1992). In frozen tissues (Wei and Giebel 1991), only some sections showed enough morphology to recognize tissue organization, but there was few cells even when liquid nitrogen was used (Soliman 1988a,b). This technique must be restricted to the identification of antigens, receptors, etc., when they were poorly preserved by other methods.…”

“…This technique must be restricted to the identification of antigens, receptors, etc., when they were poorly preserved by other methods. Moreover, this is not a routine technique in experimental or in clinical laboratories (Wei and Giebel 1991).…”