Project Summary Temporal bone research is essential for refining our understanding of the mechanisms of otologic disease; however, the difficulty in accessing this tissue, the typically long processing times, and the expense have contributed to a dwindling number of labs engaged in this work. In the 1920s, the first human temporal bone laboratory in the U.S. was established at Johns Hopkins. The collection contains approximately ~1400 processed specimens from donors with a range of otologic diseases that were obtained primarily in the 1920s through 1970s, but only a handful of studies have made use of this collection in the past 20 years. We propose to make the collection more accessible to the scientific community and to re-establish the infrastructure for harvesting, imaging, and processing new specimens with the intention of recruiting new donors from population-based studies of aging in which participants have already consented to autopsies and brain donations being performed at Johns Hopkins, as well as other clinical otological studies ongoing in the Baltimore Metro Region. We will partner with faculty specializing in biological visualization, bilingual scientific communication, and mentoring diverse scientists to develop inclusive outreach tools. We will also partner with Baltimore-based transcriptomics resources to establish protocols for performing spatial transcriptomics analysis in nonhuman primate and human temporal bones. We will approach these objectives with three Specific Aims: 1) Catalog, prioritize, and digitize existing specimens from the Johns Hopkins Human Temporal Bone Collection; 2) Develop efficient harvesting, rapid histology, and spatial transcriptomics workflows for new specimens and establish partnerships for collecting specimens during autopsies performed on participants in epidemiological studies; 3) Education/outreach activities to encourage other investigators to engage in temporal bone research. The large temporal bone collection and the inner ear illustrations of Max Brödel at Johns Hopkins contributed greatly to our early understanding of many diseases of the ear, such as the discovery of glomus bodies, the histopathologic correlates of presbycusis, and innervation of the saccular maculae. The collection has also shown enduring value, contributing to our understanding of the prevalence of a dehiscent superior semicircular canal and arachnoid granulations as well as the role of melanin in cochlear and vestibular protection. Sharing this valuable collection with other researchers while applying novel techniques in digital processing and education will accelerate advances in and knowledge of otopathology. We look forward to reinvigorating this tradition of excellence by developing a new collection, applying modern techniques, and integrating otopathology with population-based studies that include comprehensive otologic testing and functional measures.