This study demonstrates the mechanism of sensory hair fusion and its relationship to glycocalyx on the hair cells in the vestibular epithelia of gentamicin-treated guinea pigs, using a ruthenium red staining technique. 0.1 ml of a solution containing 50 mg gentamicin sulphate per 1 ml was injected once into the middle ear. After 7 days, various stages of sensory hair degeneration were observed. The glycocalyx was clearly visualized by ruthenium red. The gentamicin-induced hair damage started with a local decrease in or loss of glycocalyx from neighbouring sensory hairs, followed by formation of local attachment areas of membrane sensory hairs. These findings suggest that the glycocalyx of the sensory cell may play an important role in separating the cilia as well as maintaining the organization of the bundle. Therefore, the decrease in or loss of glycocalyx from the hair bundle of sensory cells affected by gentamicin, either directly or secondarily, can be considered as closely related to the fusion of the sensory hairs.