We measured sound-evoked vibrations at the stereociliary side of inner and outer hair cells and their surrounding supporting cells, using optical coherence tomography interferometry in living anesthetized guinea pigs. Our measurements demonstrate a gradient in frequency tuning among different cell types, going from a high best frequency at the inner hair cells to a lower one at the Hensen cells. This causes the locus of maximum inner hair cell activation to be shifted toward the apex of the cochlea as compared to the outer hair cells. These observations show that additional processing and filtering of acoustic signals occurs within the organ of Corti prior to inner hair cell excitation, thus reinstating a transformed second filter as a mechanism contributing to cochlear frequency tuning.