Leica SP5 confocal microscope for Pittsburgh Center for Kidney Research Imaging C

描述(申请人提供)：此申请是为了获得购买徕卡TCS-SP5光谱共聚焦显微镜的资金。该仪器将安装在匹兹堡大学医学院由NIH资助的P30匹兹堡肾脏研究中心的尿路上皮成像核心中。核心的功能是提供尿路的定性和定量图像分析，包括排列在肾脏肾单位和膀胱粘膜表面的特化上皮细胞。该中心为匹兹堡大学和其他机构的一大批由NIH资助的研究人员提供服务。共焦显微镜是岩心提供的一种基本成像工具，对岩心研究人员开展的研究至关重要。核心目前容纳了一个已有7年历史的徕卡TCS-SL系统，难以检测到低强度信号，无法同时捕获两个以上的荧光通道，一个低输出(1 MW)的红色He-Ne激光器，有限的激光线路范围，有限的软件能力，以及无法更新的系统架构。所要求的仪器将：1)提供无滤光片/二向色性的光路，与冷却的数字PMT结合可检测到微弱信号；2)提供4个同时检测通道；3)扩展我们现有的激发波长，使其包括405 nm、458 nm、476 nm、488 nm、496 nm、514 nm、561 nm和633 nm的激光谱线；4)提供新的软件形式，使用户能够轻松地执行FRAP、FRFT和去卷积；5)提供一种随新技术出现而升级和扩展显微镜功能的轻松途径。

DESCRIPTION (provided by applicant): 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.