LEICA TCS SPII SPECTRAL CONFOCAL MICROSCOPE

由S NIH资助的9个项目的研究者组成的主要用户组，沿着4个NIH资助的次要用户，提议购买Leica TCS SP光谱共聚焦显微镜系统。这些研究人员的工作需要最先进的共聚焦能力，而目前可用的过时的11年系统无法满足这些能力。这些主要项目需要多个激发带、用于发射的可调带宽、用于同时成像的多个通道、改进的灵敏度、真实的实时成像能力和用于真实的实时成像和三维重建的最新计算机支持。预期提出的共聚焦实验将增加我们对光感受器中的时钟调节蛋白质和基于微管的运动性（Besharse，2个项目）、细胞表面的整合素和PECAM-1的结构组织和信号传导（纽曼，2个项目）、血管性血友病抗原II的结构和功能（蒙哥马利，3个项目）、血管性血友病抗原II的结构组织和信号传导的理解。 和E-钙粘蛋白和视网膜色素上皮细胞的极性（Burke）以及牛痘病毒装配期间病毒工厂的三维组织（Traktman）。每个项目都涉及基本的细胞生物学问题，需要在NIH资助的大型项目中使用复杂的共聚焦系统。在拟议项目的三年期间，主要用户组预计将在至少80%的可用时间内使用新系统;其他NIH资助的研究人员（包括列出的次要用户）将在系统可用时访问。新仪器将在威斯康星州医学院现有的成像核心设施内安装。该成像设施由一名全职生物医学工程师进行日常监督，该工程师将负责仪器维护和实验室工作人员培训。在项目期间，新系统的使用将由一个咨询委员会控制，该委员会由主要用户组成，并由首席调查员担任主席。该机构将承担仪器的所有翻新费用和服务要求。项目结束后，NIH资助的研究人员使用的仪器的长期护理将置于现有成像核心设施的监督下。

A major user group consisting of S NIH funded investigators with 9 projects, along with four NIH funded minor users, propose to purchase a Leica TCS SP Spectral Confocal Microscopy System. The work by these investigators requires state of art confocal capabilities that are not met by an out-dated 11 year old system currently available. These major projects require multiple excitation bands, tunable band-widths for emission, multiple channels for simultaneous imaging, improved sensitivity, real- time imaging capability and state of the art computer support for real time imaging and three-dimensional reconstruction. The confocal experiments proposed are expected to increase our understanding of clock regulated proteins and microtubule based motility in photoreceptors (Besharse, 2 projects), structural organization and signalling by integrins and PECAM-1 at the cell surface (Newman, 2 projects), structure and function of von Willebrand Antigen II (Montgomery, 3 projects), structural organization of and E-cadherin and polarity of retinal pigment epithelial cells (Burke) and three dimensional organization of viral factories during vaccinia virus assembly (Traktman). Each project addresses fundamental cell biological questions requiring a sophisticated confocal system in the context of a larger NIH funded project. The major user group is expected to use the new system for at least 80% of the available time during the three years of the proposed project; other NIH funded investigators including listed minor users will have access when the system is available. The new instrument will be set up within an existing imaging core facility at Medical College of Wisconsin. This imaging facility is supervised on a day to day basis by a full-time Biomedical Engineer who will assume responsibility for instrument maintenance and training of laboratory staff. During the project period, use of the new system will be controlled by an Advisory Committee consisting of the major users and chaired by the Principal Investigator. The institution will cover all renovation costs and service requirements for the instrument. After the project period, the long term care of the instrument for use of NIH funded investigators will be placed under the supervision of the existing imaging core facility.