Deterioration of hearing is a significant problem in all age categories worldwide: the WHO estimates that one in ten people will have disabling hearing loss by 2050. Ossicular abnormalities in the middle ear can lead to conductive hearing loss and a reconstructive surgery (ossiculoplasty) is often needed, while still plagued by a high failure rate. In this project, synchrotron radiation-based X-ray dynamic micro-tomography will enable the direct visualization of the tiny and fast movements of the three auditory ossicles in entire human middle ears under a wide range of acoustic stimulation frequencies and intensities, and the ossicular chain biomechanics will be quantitatively characterized by an automated high-throughput analysis pipeline. These dynamic analyses will be completed by the description of the nano-scale arrangement of the auditory ossicles with small-angle X-ray scattering tensor tomography. Conducted on healthy and then reconstructed human middle ears with numerous configurations and surgical techniques, this project will shed light on the optimal design and positioning of prostheses as well as the optimal sculpting procedure of autologous material, globally improving ossiculoplasty success rate.