PROJECT SUMMARY/ABSTRACT Ten funded user groups at the University of South Alabama College of Medicine (USACOM) are seeking support under the Shared Instrumentation Grant Program to obtain a Zeiss LSM 980/Airyscan confocal microscope. The Zeiss system is a high-end confocal microscope with both high sensitivity spectral imaging capabilities and Airyscan, making it unique in the confocal microscopy landscape. The Zeiss LSM 980/Airyscan confocal microscope will provide spectral 2D and 3D imaging capabilities for a wide range of investigations involving live cells and tissues. In addition, Airyscan provides a unique approach to markedly increasing instrument sensitivity, such that there is little photobleaching/photodamage to samples, even during prolonged imaging experiments. The current application targets federally funded research projects that critically need these cutting-edge capabilities. These high-priority projects include ongoing studies in the areas of Pulmonary Physiology, Cellular Host Response, Biomedical Engineering, and Neuroscience. Acquisition of a Zeiss LSM 980/Airyscan system is justified to support these users: 1) our three confocal microscopes at the University of South Alabama BioImaging Core Facility (BCF) - a PerkinElmer RS3, an Andor WD spinning disk, and a Nikon A1R - are heavily used and two of the units are vintage (14 and 9 years for the Perkin Elmer and Nikon systems, respectively) and in increasing disrepair; and 2) the current imaging systems simply cannot provide key capabilities required by NIH-funded investigators – specifically high sensitivity spectral imaging and sustained gentle imaging with little photobleaching/photodamage. Critical equipment and personnel are in place, and we successfully assembled and tested both Zeiss and Nikon systems during recent equipment demonstrations. Acquiring a Zeiss LSM 980/Airyscan systems will dramatically extend our ability to perform simultaneous acquisition of multiple fluorescence signals in live cells and tissues – in spectral or Airyscan modes – and will push ongoing NIH-funded research projects beyond their current limitations. In the long range, including the Zeiss LSM 980/Airyscan system within our existing BioImaging Core Facility at the USACOM will rapidly advance existing projects and allow pursuit of new experimental directions. This will be an invaluable tool for seeding new collaborative and translational initiatives among our research faculty.