Hearing and balance rely on the ability of hair cells in the inner ear to sense miniscule mechanical stimuli. In each cell, sound or acceleration deflects the mechanosensitive hair bundle, a tuft of rigid stereocilia protruding from the cell's apical surface. By altering the tension in gating springs linked to mechanically sensitive transduction channels, this deflection changes the channels' open probability and elicits an electrical response. To detect weak stimuli despite energy losses caused by viscous dissipation, a hair cell can use active hair-bundle movement to amplify its mechanical inputs. This amplificatory process also yields spontaneous bundle oscillations. Using a displacement-clamp system to measure the mechanical properties of individual hair bundles from the bullfrog's ear, we found that an oscillatory bundle displays negative slope stiffness at the heart of its region of mechanosensitivity. Offsetting the hair bundle's position activates an adaptation process that shifts the region of negative stiffness along the displacement axis. Modeling indicates that the interplay between negative bundle stiffness and the motor responsible for mechanical adaptation produces bundle oscillation similar to that observed. Just as the negative resistance of electrically excitable cells and of tunnel diodes can be embedded in a biasing circuit to amplify electrical signals, negative stiffness can be harnessed to amplify mechanical stimuli in the ear. U niquely among sensory receptors, the hair cells in the ears of tetrapod vertebrates use mechanical feedback to amplify their inputs. Mechanical amplification endows these animals with both exquisite auditory sensitivity and sharp frequency discrimination (reviewed in refs. 1-3). More specifically, by providing energy to compensate for that lost to viscous dissipation in the ear's fluids, the amplifier permits each receptor organ to act as a highly tuned resonator (4). The ear's active process is characterized by metabolic vulnerability, an intimation of powered amplification, and by the spontaneous emission of sounds in a quiet environment, a sign of excess feedback gain.The mechanism by which mechanical energy is produced by the vertebrate inner ear remains uncertain. The active process of mammals is thought to involve electromotility, a voltage-induced change in the length of the outer hair cell (reviewed in refs. 5-7). In nonmammalian tetrapods, whose hair cells lack electromotility, amplification apparently involves active hair-bundle movements. In response to abrupt deflections, active hair bundles can twitch, performing work against an external load (8). Active bundles can exhibit spontaneous oscillations (9-11) that may underlie spontaneous otoacoustic emissions. Finally, spontaneously oscillatory hair bundles can amplify periodic mechanical stimuli (12). In the present work, we have examined the mechanical properties of active hair bundles under displacement-clamp conditions. By so doing, we have identified a mechanism by which hair bundles produce oscilla...