Figure 13.

Material prepared originally for LSFM can be rehydrated and the basilar membrane (with organ of Corti complex) dissected free for further confocal examination. The basilar membrane as it appears in a light sheet image before rehydration (a) and in a confocal image after dissection and flat-mounting (b). Panel b is a single image snap-shot taken prior to examination at 63x oil (c-g). In the higher magnification images there was no observable membrane disruption; nuclei of both inner and outer hair cells (IHC, OHC) were evident (arrows in c) as were cuticular plates of inner hair cells (arrows in d). Deconvolved and enlarged images (e and f) show outer and inner hair cells at two focal levels (asterisk at upper left marks same outer hair cell in e and f). Here notice stereocilia of both inner and outer hair cells was preserved (hollow arrowheads in e and f). In g, the stack has been re-sliced to show the row of inner hair cells, their soma, nuclei, cuticular palates and stereocilia (hollow arrowheads point to same inner hair cells in e and g). Scale in c same for d; in e same for f and g.