A simple procedure of vibratome sectioning the organ of Corti, followed by immunohistochemistry and confocal microscopy is described. This procedure allows for improved preservation of the fine cytoarchitecture of the mammalian organ of Corti, and consequently allows for accurate quantification of cell types.
The overall goal of the following experiment is to image the organ of corti with maximal preservation of cellular architecture. This is achieved by sectioning, dissected and fixed inner ears with a vibrato to obtain thick sections while minimizing the number of cut surfaces per cochlear. The Vibram sections are then stained by immunohistochemistry, which allows the identification of various cell types and cellular structures in the organ of corti.
Next, the sections are mounted and imaged by confocal microscopy in order to obtain high resolution images that define Z positions Throughout each thick Vibram section results are obtained that allow for the confocal microscopy of the cells and structures in the organ of corti with minimal disruption by the sectioning process. The main advantage of this technique over existing methods such as paraffin and cryosectioning, is that physical damage to the cochlea is minimized, allowing better preservation of the cellular architecture of the organ of corde In order to isolate the inner ear. First, decapitate a euthanized adult mouse using a razor blade.
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