SHARED CONFOCAL MICROSCOPY FACILITY

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DESCRIPTION (provided by applicant): This revised application requests a confocal microscopy system to serve a group of NIH-funded investigators in the Departments of Neurology and Molecular and Cellular Pharmacology, and the Cerebral Vascular Disease Research Center (CVDRC), at the University of Miami School of Medicine. Six NIH-funded program application areas are described. In two programs, confocal microscopy is applied to whole-animal brain-ischemia models in vivo to obtain high-resolution dynamic imaging of vascular wall morphology, plasma and red cell flow, leukocyte adherence, and thrombus development and lysis. Several programs also utilize confocal slide-microscopy for single- and multiple-label immunohistochemistry. The requested system will also be used for confocal imaging of the organotypic hippocampal slice preparation and for livecell imaging. The in vivo applications require that the confocal system be located within our animalexperimentation laboratories, which contain instrumentation and resources for microsurgery, physiological monitoring, and laser-based photothrombosis. The confocal instrument will be integrated into the CVDRC's Image-Analysis Core facility, and software developed to co-map confocal images into a multimodal image database for correlative regional analysis of multi-label immunohistochemistry with blood-flow and metabolism autoradiography, histopathology, and in situ hybridization data sets. The six NIH-funded program application areas include: 1) The mechanisms responsible for laser-induced lytic recanalization of platelet thrombi induced by photothrombotic arterial occlusion, and their relevance to reperfusion injury in the brain. 2) Focal cerebral ischemia: mechanisms of albumin-neuroprotection and hyperthermic exacerbation. 3) Synaptic alterations and neuroplasticity in ischemic brain. 4) The consequences of chronic cerebrovascular occlusions. 5) Ischemic preconditioning and mitochondrial mechanisms. 6) Programmed death signaling pathways in cardiac ischemia and reperfusion, and the therapeutic role of angiogenesis.