ABSTRACT: MICROSCOPY AND IMAGE ANALYSIS CORE The proposed COBRE will establish a center of excellence at the University of Maine (UMaine) focusing on the impacts of extracellular signaling on cell behavior. The five interconnected projects in this proposal will elucidate the intricacies of extracellular signaling, which will require sophisticated and dynamic analyses of cellular structure in vivo. In addition, all five of the proposed research projects are structured around longitudinal designs that require higher throughput imaging of live systems over a longer period of time. The five projects will also incorporate novel, unbiased image analysis paradigms that will increase rigor and reproducibility. To address these needs, the proposed COBRE Microscopy and Image Analysis Core (MIAC) has two specific aims to provide training and access to (1) state-of-the art, high resolution live imaging services and (2) experiment planning, image acquisition and image analysis services as well as data storage and management. MIAC will provide training and access to state-of-the-art, high resolution live imaging services, including a recently-acquired inverted Leica confocal/lightsheet microscope, an upright Zeiss Airyscan2, and an Olympus FV-1000 confocal. In addition, funding through the NIH COBRE program will allow UMaine to acquire an inverted Zeiss LSM900 with Airyscan2 confocal microscope designed for live imaging of infectious disease, which will be a unique capacity in the state. MIAC will provide wraparound services from experimental planning to expert data management and image analysis. These services are critical for rigor, reproducibility and maximizing the impact of microscopy equipment on the proposed research projects and for other users. The MIAC core will provide one-on-one mentorship of users in planning, executing, analyzing, and storing their data. This will include instruction in the theory and practice of microscopy, as well as ongoing support for microscope troubleshooting, image analysis with Zen, ImageJ, Matlab, Imaris and Aivia software packages. The MIAC core is innovative in that it will leverage existing engineering, physics and biochemistry expertise in imaging and analysis to guide projects and exploit the power of our microscopes and analysis software from start to finish. The establishment of a Microscopy and Image Analysis Core at UMaine will fulfill important gaps in biomedical infrastructure in the state and catalyze important new discoveries in how cell behavior is governed by extracellular signaling in multiple contexts.