Confocal scanned imaging light microscopes became commercially available in 1987. There was instant excitement among the biological community. Details of cellular and subcellular structures labelled with fluorescent dyes or reflective gold deep in tissues are visualized with great clarity. Scattered light from out-of-focus tissue above and below the plane of focus is almost completely eliminated. Three dimensional reconstructions of the tissue can be developed from a series of optical images acquired as the plane-of-focus is scanned through the tissue. Non-destructive high-resolution images of living tissues labeled with voltage- and ion-sensitive dyes follow dynamic processes of cell and organelle function. Many investigations of spatial relationships which were difficult and immensely time consuming are now relatively straightforward. We are now in the second generation of instrument development. Experimental limitations of the technique are well understood. Instrumental versatility is increased. Different instruments are best suited to different investigations. Investigators now have a fairly clear understanding of which of their scientific goals can be realized more effectively and of what questions which had not been experimentally practical are now possible. This Shared Instrumentation Grant application is submitted by a group of investigators with extensive PHS grant funding a strong commitment to the application of front-edge techniques to their studies, and a realization that installation of a confocal imaging facility at the University of Cincinnati will stimulate the research effort well beyond the goals of their specific projects. The instrument proposed is well-suited for all of the investigator's projects. By sharing an instrument facility optimized for their needs an instrument that would be prohibitively expensive for a single investigator becomes accessible. Investigators are in the Departments of Anatomy & Cell Biology and of Molecular Genetics, Biochemistry & Microbiology. The faculty in these departments are among the most substantially funded in medical colleges in the United States. Projects of the investigators include studies of receptor neuron stimulus selectivity, fibroblast differences in pulmonary fibrosis, structure and dynamics of cytoskeltal elements, plasticity in the neuroendocrine system, organization of cortical neural networks, metabolic signaling by movement of intracellular lipids, and virus reactivation in nerve cells. The investigators are widely recognized for work using imaging techniques to advance understanding of structure-function relationships and have introduced major advances in research techniques. This proposal is for funds for a laser scanning confocal microscope and image analysis instrument which will advance their research efforts. The projects of each of the investigators are high-priority, health-related scientific studies in the fields of neuroscience, and cell and molecular biology.
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