Effects of Fixation Methods on Cochlear Staining in Mice
WEI Gege
QIU Shiwei
XIONG Wei
YUAN Yongyi
NIE Huiyi
DAI Pu
Abstract：Purpose To report effects of fixation techniques on demonstration of general structures and cell mor-phology in the cochlea with preservation of cellular structures and intracellular organelles,to improve current protocols for best immunostaining results.Method One-month-old wild-type C57BL6J mice with normal hearing(n=9)were ran-domly divided into three groups with three mice in each group.Under general anesthesia,animals were perfused with sa-line + 4%paraformaldehyde(PFA)(Group 1)or 4%PFA directly into the heart(Group 2),or decapitated with the co-chlea harvested and directly perfused with 4%PFA.The fixed cochlea was cryosectiond and stained for immunofluores-cence,followed by laser confocal scanning microscopy imaging at low and ultra-high magnification.Results Structures of hair cells and auditory nerves were clearly revealed by all three fixation protocols with similar qualities(P>0.05).However,the structure of stria vascularis was better preserved in group 1 than the other two groups.Conclusions The three fixation methods in this study are all adequate for morphology studies of the organ of Corti and spiral ganglia,while saline + 4%PFA perfusion may be preferable for studying ultrastructure of the stria vascularis.
Keywords：cardiac perfusioncochlear perfusioncorti's organstria vascularisspiral ganglion
Publication Date：2024-02-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 141-145 )