Dissection technique for cochleas prepared for scanning electron microscopy

Scanning electron microscopy (SEM) permits a three-dimensional study of the surface morphology of the organ of Corti that is very useful in evaluating the condition of the apical end of the hair cells and the stereocilia. However, some laboratories have experienced problems with curling of the basilar membrane during critical point drying of cochlear specimens prepared for SEM evaluation using the Murakami or osmium thiocarbohydrazide-procedures. This curling of the basilar membrane can obstruct the view of the reticular lamina and the ciliary ends of the hair cells. We have used a dissection method, referred to as the anchor technique, to overcome basilar membrane curling. This technique removes all the structures above the reticular lamina but leaves the basilar membrane attached to the spiral ligament and the lateral bone to which the spiral ligament is anchored. Individual cochlear turns are dissected in this manner and mounted on the same examination stub for SEM evaluation. Maintenance of the lateral attachment of the basilar membrane requires additional dissection time but eliminates the problem of curling during critical point drying. An additional benefit is that mounting the individual turns on the same examination stub facilitates evaluation and photomicroscopy of the surface morphology. The anchor technique has been used successfully on the guinea pig and should be appropriate for most mammalian cochleas.