ShEEP Request for Laser Scanning Confocal Microscopy Acquisition and Analysis System

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This proposal is for a shared Laser Scanning Confocal Microscopy Acquisition and Analysis System to allow a Major User group (six Merit Award funded PIs) as well as other VA researchers at the VA Ann Arbor Health Care System (VAAAHS) to conduct high-resolution analysis in both living and preserved tissues. Research focuses on cellular pathogenesis of diseases that preferentially affect veteran populations. Studies include firearm triggered loss of hearing and balance, Deployment-Related Chronic Lung Disease, PTSD, COPD, CNS infections and subset of neurodegenerative disorders, certain types of cancer (tumor immunotherapy). These diseases are characterized by specific, and often subtle, micro-anatomic tissue changes and a detailed identification and assessment at the molecular level is critical to execute studies, propose new treatment opportunities and subsequently test their effectiveness. This need is not being adequately met. The requested system is composed of Leica SP-8 inverted laser scanning confocal microscope with a Tokia HIT stage-top incubation system for live cell imaging. It will have multiple lasers and high capability detectors with adjustable channels, rapid acquisition capacity and up to 120 nm resolution. Air, water, oil and glycerol immersion objectives with varying working distances and numerical apertures will match the needs of our diverse user group for their variety of specimens and assessments. Software on both acquisition and image analysis workstations will provide for enhanced imaging, visualization, co-localization and rotation as well as 2-D and 3- D analysis and measurement. FRET and FRAP “wizards” will allow these applications. Introducing new capabilities will enhance current studies by providing new data and improved data collection and analyses as well as increasing efficiency and throughput. It will extend current studies into new directions by allowing identifications and assessments that would not otherwise be possible, for example, by allowing new multiple co-labeling of molecules and cellular elements deep into specimens or by allowing real-time dynamic live cell imaging, FRET and FRAM analyses. It will also create new capabilities aiding future grant applications of our faculty and new learning opportunities for trainees at the VAAAHS site. The requested system will enhance and extend VAAAHS research on disorders effecting our veteran population towards developing therapeutic interventions to ultimately benefit our patients.