Leica Stellaris 8 Confocal Microscope System

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7. Project Summary/Abstract Our ability to understand cellular function in health and disease from the macro to micro to nano and even to the atomic scale depend on the capabilities of state-of-the-art advanced instrumentation. The NIH funded projects described in this S10 application require enhanced imaging resolution and multi-spectral capabilities for their fluorescence microscopy applications to advance to the next level. A Leica Stellaris 8 laser scanning confocal microscope is requested and will be located in the UMKC Confocal and Multiphoton Microscopy Core Facility. This instrument will replace and upgrade our current Leica TCS II Sp5 confocal microscope that operates as a shared resource to support the research programs of a core group of NIH funded major users as well as minor and other users in the UMKC Schools of Dentistry, Pharmacy, Medicine, Biological Sciences, Computing and Engineering, and the nearby Children’s Mercy Hospital. The user group has a diverse research emphasis on musculoskeletal research, neural/vision research, developmental biology, craniofacial and dental research, molecular biology, yeast genetics, neurodegeneration, aging, cancer and drug delivery, materials science, and even petrochemical research. To accommodate the wide-ranging project applications, the instrument will be configured for confocal microscopy on fixed cell and tissue specimens, and for live cell imaging, multispectral imaging, fluorescence recovery after photobleaching (FRAP), optical sectioning/3D imaging, and reflection imaging. Three of the projects employing this technology are examining mechanisms of signaling crosstalk between muscle and bone that may regulate age related osteoporosis and sarcopenia. The projects are using live cell imaging to monitor mitochondrial function and extracellular vesicle-mediated muscle-bone communication and are using 3D confocal imaging to assess the cellular and structural organization of bone in aged gain and loss of function mouse models with and without exercise. One project is using live cell imaging to examine mechanisms controlling calcium signaling to prevent neurodegeneration in early stage glaucoma and two projects are developing drug delivery and tissue engineering approaches to treat age related macular degeneration. Other projects are focused on molecular and cellular mechanisms of cleft palate formation, heart contractile function and calcium signaling, developmental jaw morphogenesis, single cell and spatial genomics in pediatric bone tissues, finite element and fluid dynamic modeling of osteocyte mechanotransduction and nanoparticle systems for drug delivery in cancer and fibrosis. The Stellaris 8 confocal system will accelerate the progress of these projects by providing an exceptional level of resolution and sensitivity that is needed for 3D cell and tissue imaging and live cell imaging in mineralized and other tissues. The multi-spectral imaging capabilities of the Stellaris 8, the versatility of its excitation and detection capabilities, its enhanced sensitivity and its TauSense/ fluorescence lifetime technology are critical to advance this research to the next level, and will allow for expansion of our capabilities for multiplexing, 3D spatial mapping and cleared tissue imaging. Acquisition of this technology will accelerate discoveries in musculoskeletal health, development, aging, vision, dental restorations, and drug delivery.