Fig. 4.

Celloidin removal with acetone followed by immunostaining. Reissner’s membrane (black arrows) stains in many panels (nonspecific staining). Supporting cells (asterisks) are diffusely brown in many panels. With Na⁺,K⁺-ATPase antibody, stria (S) starts to stain appropriately, but there is a great deal of background staining. Celloidin (gray arrows) can still be seen in some panels. Spiral ligament (arrowheads) shows background staining. Tubulin panel shows appropriate staining in root cells (R) and in a portion of pillar cells (P), but background staining is also seen. Neurofilament antibody stains nerve fibers (N) below inner hair cell area, but it also spuriously stains spiral ligament (arrowhead). Calibration bar — 50 µm.