It is well known that the migration of epidermal cells of the tympanic membrane has important roles in healing process of the tympanic membrane perforation and self cleansing mechanism of the external auditory canal. However, the migration mechanism of epidermal cells is known a little. Recently, there are some reports that epidermal cells of the tympanic membrane have contractile protein which is related to motility of cells and provides contraction force to cells in cellular migration. Authors performed this study for observing contractile protein in epidermal cells of the tympanic membrane. Guinea pigs of both sex weighting 250 to 300 gm were used. We fixed the tympanic membranes with instillation of fixatives(Histochoice, Amresco) into the external canal and bulla, and separated the tympanic membrane. Immunohistochemical stainings for the contractile protein in the epidermal cells were observed under microscope. On immunohistochemical staining, the tympanic membrane showed homogenous and red staining but no staining in negative control. Radiating fibers directing from handle of malleus to annulus were also noted in the lamina propria. Strong and homogenous staining was observed in the cytoplasm of basal cells but not in the nucleus. Prickle cells and granular cells showed very weak staining comparing with the basal cells, and immunostaining was more prominent in marginal portion than in perinuclear area. Keratin layer was not stained. This findings suggested that epidermal cells of the tympanic membrane have contractile protein which could be related to the epidermal migration.