The middle ear is too complex a system for its function to be fully understood with simple descriptive models. Realistic mathematical models must be used in which structural elements are represented by geometrically correct three-dimensional (3D) models with correct physical parameters and boundary conditions. In the past, the choice of boundary conditions could not be based on experimental evidence as no clear-cut data were available. We have, therefore, studied the deformation of the tympanic membrane (TM) at its boundaries using X-ray microscopic computed tomography in human and gerbil while static pressure was applied to the ear canal. The 3D models of the TM and its bony attachments were carefully made and used to measure the deformation of the TM with focus on the periphery and the manubrium attachment. For the pars flaccida of the gerbil, the boundary condition can, for the most part, be described as simply supported. For the human pars flaccida, the situation is more complicated: superiorly, the membrane contacts the underlying bone more and more when pushed further inward, and it gradually detaches from the wall when sucked outward. In gerbil, the attachment of the TM to the manubrium can be described as simply supported. In human, the manubrium is attached underneath the TM via the plica mallearis and the contact of the TM with the bone is indirect. For both human and gerbil, a simple boundary condition for the peripheral edge of the pars tensa is not appropriate due to the intricate structure at the edge: the TM thickens rapidly before continuing into the annulus fibrosis which finally makes contact with the bone.