This request is for funds to purchase a Leica TCS-SP5 spectral confocal microscope. This instrument will be housed within the Urinary Tract Epithelial Imaging Core of the NIH-funded P30 Pittsburgh Center for Kidney Research at the University of Pittsburgh School of Medicine. The function of the core is to provide qualitative and quantitative image analysis of the urinary tract, including the specialized epithelial cells that line the nephron of the kidney and mucosal surface of the bladder. The core serves a large group of NIH- funded investigators at the University of Pittsburgh and at other institutions. Confocal microscopy is an essential imaging tool provided by the core and is crucial to the research preformed by core investigators. The core currently houses a >7-year old Leica TCS-SL system with difficulty detecting low intensity signals, inability to capture more than two channels of fluorescence at one time, a red He-Ne laser with low output (1 mW), a restricted range of laser lines, limited software capabilities, and a system architecture that cannot be updated. The requested instrument will: 1) provide a light path with no filters/dichroics, which when coupled with cooled digital PMTs can detect weak signals;2) provide 4 channels of simultaneous detection;3) extend our available excitation wavelengths to include laser lines at 405nm, 458 nm, 476 nm, 488 nm, 496 nm, 514 nm, 561 nm, and 633 nm;4) provide new software modalities that allow users to easily perform FRAP, FRET, and deconvolution;5) provide an easy path to upgrade and expand the capabilities of the microscope as new technologies become available.
