Immunohistochemical and morphological studies on the human fetal cochlea: a comparative view on methods.
The preservation of morphology and antigenicity can vary uncontrollably with human fetuses since these rely heavily on immediate fixation of the temporal bone following spontaneous abortion. Once good fixation is established, there is the question of the approach taken for morphologic and immunohistochemical studies. To achieve maximal preservation for the purpose of studying normal and pathologic fetal cochleae, commonly used preparation methods for analyzing the cochlea were reviewed and compared for both immunohistochemical and morphologic studies. Cochleae obtained after spontaneous abortion ranged from the 9th gestational week to birth. Four different methods were compared for morphologic study: the block surface method; a microslicing technique; paraffin; and celloidin sectioning. For immunohistochemical study, three methods were compared: pre-embedding; paraffin; and frozen sectioning. For morphologic preservation, the block surface method gave best overall results, showing good representation of the fetal cochlea for surface preparation, light, and electron microscopy. Celloidin sectioning was also found to show good light microscopic results for both the middle and inner ear. To achieve optimal results, preservation quality, fixation procedures, and antibody all contribute to the efficacy of a methods choice.
Fish
J H
JH
Center for Inner Ear Biology, Department of Otorhinolaryngology, University of Innsbruck, Austria.
Scholtz
A W
AW
Hussl
B
B
Kammen-Jolly
K
K
Ichiki
H
H
Kreczy
A
A
Schrott-Fischer
A
A
D000906:Antibodies; D003051:Cochlea / Q000737:chemistry* / Q000196:embryology*; D003101:Collodion; D000072259:Connexin 26; D017630:Connexins / Q000032:analysis* / Q000276:immunology; D005629:Frozen Sections; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016612:Paraffin Embedding; D014014:Tissue Adhesives; D014404:Tubulin / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D017630:Connexins; D014014:Tissue Adhesives; D014404:Tubulin; D000072259:Connexin 26; D003101:Collodion