This proposal requests funds to purchase a laser scanning confocal microscope system with FRET and TIRF microscopic capabilities. The instrument will be housed within the imaging facility of Dalton Cardiovascular Research Center (DCRC) at the University of Missouri. While several modern fluorescent microscopes are available for general use, we do not have a confocal microscope at this site. There is a high demand placed on the fee-for-service access to the single confocal microscope available at this university creating shortage of large time blocks necessary for conducting live cell/tissue imaging experiments. In view of the time- sensitive nature of our experiments and difficulty in transporting live cells/ tissue to this location, the addition of a confocal microscope to the imaging core at DCRC would greatly enhance a diverse array of ongoing NIH-funded research programs. As detailed in this proposal, an advisory committee was formed to identify a core group of users, determine the specific needs of DCRC and to choose the appropriate instrumentation to accomplish the goals of our research groups. Following extensive consultation and analysis of various competing confocal systems, the Olympus Fluoview 1000 has been identified as the ideal system. This system can be modified to integrate FRET and TIRF capabilities with patch-clamp techniques to serve the needs of investigators applying innovative fluorescent microscopy and nanotechnology approaches to central questions in membrane physiology such as exocytosis/ endocytosis, ion channel gating and receptor- mediated signal transduction. In addition, this confocal system will also support a second group of investigators studying subcellular localization of glutamate and GABA receptors and neuronal pattern generation in the central nuclei involved in blood pressure regulation. The addition of the proposed confocal microscope will not only maximize the impact of our existing research programs but also will help in recruiting new, talented faculty to join the already successful group at our Institution. ? ? ?
