PROJECT SUMMARY ? Overall. This application proposes the founding of a Tissue Mapping Center (TMC) for the human eye and pancreas within the Human BioMolecular Atlas Program (HuBMAP). The mission of the proposed TMC is to build a platform of integrated technologies for imaging and molecular analysis that enables the construction of comprehensive 3-dimensional molecular atlases of human eye and pancreas. This TMC will leverage the unique resources of the Mass Spectrometry Research Center and the National Research Resource for Imaging Mass Spectrometry at Vanderbilt University, the world-class clinical environment of the Vanderbilt University Medical Center, the distinguished expertise of Organ Specific Project team leaders in their respective fields and their vast experience in organ procurement and management, and the advanced biocomputational infrastructure available to the TMC through the Data Analysis Core laboratories at Vanderbilt University and the Delft University of Technology, Netherlands to create a capability to molecularly characterize human tissues in 3-dimensions at a level of understanding unrivaled by current technologies. The innovative aspects of the proposed TMC are (1) the integration of imaging mass spectrometry, co-detection by indexing (CODEX), stained microscopy, and autofluorescence microscopy into 3-dimensional molecular atlases, (2) whole organ imaging of the eye, and (3) multi-organ atlases from the same donors through a proposed collaboration with the currently funded University of Florida TMC. The application of the proposed platform to organ-specific projects in pancreas and eye will provide a new paradigm of understanding the normal state of these organs across vast scales, both molecular (e.g., lipids, metabolites, and proteins) and spatial (e.g., whole organs to single cells). Furthermore, the involvement of these organs in diabetes, along with organs currently funded by HuBMAP such as kidney and vasculature, will lay an important foundation for future studies striving to understand the progression of diabetes. As a HuBMAP participant, the molecular atlases produced by this TMC will be disseminated to collaborators to generate new hypotheses regarding the function of these important organ systems, enabling new insight into human health and disease. This multidisciplinary effort requires the creation and integration of three capabilities to 1) procure and manage human tissue specimens, 2) determine and mitigate non-biological pre-analytical factors, and 3) acquire, process, and disseminate multimodal 3-dimensional imaging and large-scale omics data.