Project Summary/Abstract The goal of this ORD Shared Equipment Evaluation Program (ShEEP) proposal from the Saint Louis VA Healthcare System is to obtain a high content confocal microscopy imaging system capable of obtaining and processing high resolution images of multiple samples, including live cells. The system will be shared by multiple VA-funded investigators and their colleagues at two affiliated universities Saint Louis University School of Medicine and Washington University School of Medicine. Both affiliates have substantial research enterprises connected to the Saint Louis VA. The instrument is designed to support and enhance the research endeavors of a large and diverse group of VA-funded scientists. The instrumentation requested, is a Molecular Devices ImageXpress Micro Confocal High-Content Imaging System and accompanying analysis software that includes multiple application modules pre- loaded onto an analysis workstation designed to support the imaging and data processing needs for a wide variety of investigations. The proposed system is configured for spinning disc laser confocal microscopy to accommodate rapid imaging at multiple fluorescence wavelengths, water immersion objectives that permit increased depth of penetration for thicker samples at reduced exposure times, environmental control for imaging of live samples and phase contrast for imaging of unstained cells. In addition to the need for high content imaging and analysis, a common theme among the investigators supporting this proposal is the need to image and analyze organoids. These adult stem cell-derived clusters of cells recapitulate many of the features from the tissue from which they were obtained and therefore have now been adopted in multiple lines of investigation from personalized cancer chemotherapy to microbial pathogenesis studies. Multiple projects in this proposal relating cancer biology, rheumatology, nephrology, cardiology and infectious diseases are designed around the imaging of organoids as a platform for basic discovery and translation to therapies that would benefit our patients at the VA.

Project narrative Developing new therapeutics and vaccines that support the health of our Veterans requires state-of- the-art tools. This initiative is centered on the need for advanced imaging and analysis platforms that can support multiple lines of investigation critical to basic research discovery and translational development of therapies and preventative strategies that enhance the health and welfare of our Veterans. The instrument is intended to support multiple projects by VA-funded principal investigators and facilitate a range of basic studies to advance our understanding of diseases important to Veterans. This shared instrumentation will also serve as an important focal point for collaborative efforts with VA investigators.