In this proposal, funds are requested from the National Center of Research Resources for purchase of a Leica TCS SP5 spectral confocal microscope equipped for fixed and live cell imaging to advance both the quality and expedience of scientific studies performed at the University of Pennsylvania. A core facility will be developed for the SP5 confocal instrument to be used regularly by seven primary investigators for (1) confirmation of gene delivery for cystic fibrosis and asthma in parallel with effects on cytoarchitecture and physiological responses, (2) more precise indices of hypoxia, oxidative damage and endoplasmic reticulum injury in obstructive sleep apnea, (3) in vivo neuronal activity in C. elegans to identify mechanisms of sleep, (4) z-stacks to firmly establish intracellular localization in distortion-free 3D images for studies in models of schizophrenia, and (5) elucidation of alterations in cytoskeleton and neuronal synapses in aging of sleep/wake circuits. The machine would also be made available to all NIH-funded principal investigators at the University of Pennsylvania, many of whom experience delays in NIH projects because of wait times for confocals. We have worked out a shared instrument operating system that will be overseen by the Penn Strategic Core Facility Advisory Committee that will ensure fair use times for both major user investigators and others on campus. We envision numerous productive collaborations to develop through this shared instrument usage. There is strong institutional commitment to maintain this equipment for extended time. Relevance: This instrument will enable us to more precisely define the molecular interactions in many disease processes with overriding goals to identify mechanisms of disease and then develop novel therapies for a diverse collection of diseases including sleep apnea, asthma, pulmonary hypertension, stroke, cystic fibrosis, atherosclerosis and cognitive and sleep disorders of aging.
The confocal would be used to enhance translational science to better identify molecular mechanisms underlying numerous disease processes including cystic fibrosis, pulmonary hypertension, asthma, sleep apnea, atherosclerosis, stroke, schizophrenia and sleep disorders. Elucidation of the molecular mechanisms of these diseases and conditions will be used directly to develop newer more effective therapies for these significant medical disorders, and others.
