LEICA TCS SPII SPECTRAL CONFOCAL MICROSCOPE

一个由S资助的9个项目的研究人员组成的主要用户群体，以及4个国家卫生研究院资助的次要用户，提议购买徕卡TCS SP光谱共聚焦显微镜系统。这些调查人员的工作需要最先进的共焦能力，而目前可用的过时的11年历史的系统无法满足这一能力。这些重大项目需要多个激发带、发射带宽可调、同时成像的多个通道、提高的灵敏度、实时成像能力以及用于实时成像和三维重建的最先进的计算机支持。建议的共聚焦实验有望增加我们对光感受器中时钟调控的蛋白质和基于微管的运动性(Besharse，2个项目)，整合素和PECAM-1在细胞表面的结构组织和信号传递(Newman，2个项目)，von Willebrand抗原II的结构和功能(Montgomery，3个项目)，视网膜色素上皮细胞(Burke)的结构组织和E-钙粘素与极性(Burke)，以及痘苗病毒组装过程中病毒工厂的三维组织(Traktman)的了解。每个项目都解决了基本的细胞生物学问题，在NIH资助的一个更大的项目的背景下，需要一个复杂的共聚焦系统。在拟议项目的三年期间，主要用户群体预计将使用新系统至少80%的可用时间；当该系统可用时，包括列出的次要用户在内的其他国家卫生研究院资助的调查人员将可以使用该系统。新仪器将安装在威斯康星医学院现有的成像核心设施内。该成像设施由一名全职生物医学工程师负责日常监督，负责仪器维护和实验室工作人员的培训。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，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。在项目期结束后，NIH资助的调查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之下。

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.