Funds are requested to purchase a Yokogawa CSU-W1 spinning disk confocal system coupled to a Nikon Eclipse NI-E upright microscope with Andor EM CCD detectors. This instrument will replace a Noran/Prairie Technology Scanning confocal microscope, which was purchased in 1995. The new instrument will be housed in the Neuroscience Imaging/Physiology Core Facility supported by the NIGMS/NCRR Center of Biomedical Research Excellence (COBRE) in Neuroscience grant (P30 RR032135/P30 GM103498, 7/1/2011-6/30/2016). The Neuroscience COBRE Imaging/Physiology Core is administered by a full-time, highly trained microscopist. Acquisition of the Yokogawa CSU W-1, a modern, modular, and actively supported system, would greatly enhance the live imaging capabilities of the Core, expanding the high-speed, high-resolution microscopy resources available to investigators within the College of Medicine and in other Colleges on the UVM campus. These investigators routinely record optical measurements of fast, localized intracellular events, such as calcium transients from living cells and tissues. The Yokogawa spinning disk will significantly improve the capabilities of the Core to meet the emerging needs of the present Core investigators, while providing the opportunity for additional funded and new investigators to expand the scope of their research programs. The Yokogawa system will use an upright rather an inverted microscope platform, allowing better access to intact thin tissues like isolated vessels or gut, bladder or urothelial strips by employing high numerical aperture physiology dipping lenses. The Yokogawa system can simultaneously record two low light signals with dual 512x512 EM CCD cameras at 52 fps (frames per second), a rate well above our other instrumentation. Higher resolution (2560X2160) imaging for brighter samples can be achieved with the Zyla sCMOS camera at 100 fps. These cameras allow great flexibility for the multiple sample types encountered in our multi-user facility. In addition, the Yokogawa system will have multiple laser lines, greatly broadening its experimental capacity by allowing simultaneous measurements of multiple fluorescent signals from defined cells. The COBRE Imaging/Physiology Core has a long history of supporting live imaging research and publication, but its instrumentation for high speed acquisition is obsolete, and can no longer meet current research demands.
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