Zeiss 510/META Confocal Microscope Plus Live Cell Chamber

描述（由申请人提供）：本申请要求使用一台带有环境腔的蔡司510/META激光扫描共聚焦显微镜来取代已有11年历史的蔡司410显微镜，该显微镜是NIH NIDDK核心开发中心（R24, DK064388，“霍普金斯基础科学消化疾病核心中心”，PI， M. Donowitz， M.D.）的成像核心（核心主任O. Kovbasnjuk博士，是本申请的PI）的一部分。蔡司510/META显微镜将允许我们提供最先进的荧光成像能力，5个主要的相互作用的研究人员，谁是由NIH NIDDK核心发展中心的支持，谁是密切的相互作用。同样的核心中心也支持6个次要用户，他们的实验室在罗斯研究大楼，那里有GI核心中心和蔡司共聚焦。除了作为美国国立卫生研究院资助的核心中心的一部分，510/META将成为其他需要共聚焦环境室的JHUSOM科学家的资源，并将以付费服务的方式提供给霍普金斯主要研究机构（罗斯大楼）的科学家。410不再满足用户的需求，而510/META可以满足用户的需求。这是由于1)蔡司410 PMT的恶化和针孔加上其过时的软件和脆弱的计算机支持，所以一旦当前的计算机出现故障，整个系统将无法正常工作。2)缺乏FRET应用所需的光谱扫描能力。3)由于比410更精确的像素对齐，需要广泛使用510上精度更高的FRET和FRAP。蔡司510/META的特点是完成多样化的生物医学项目所必需的，而这些特点是蔡司410所不具备的，包括：多跟踪技术，用于速度和精确的像素定位；蔡司LSM510和Axiovision软件，用于快速信号处理和分析；还有一个活细胞室，里面有温度和二氧化碳控制，需要长时间的活细胞成像。JHUSOM Imaging Core无法为R24中心提出的生理研究提供足够的时间。这阻碍了提出的FRET研究和过夜恒温室研究。重要的是，在JHUSOM按服务收费的仪器中没有配备活细胞室。蔡司510/META将安装在NIH NIDDK核心中心成像核心设施，这是大多数用户的中心位置。预计每周40小时的75%将由5个主要用户和6个次要用户使用，剩下的25%，即每周10-15个额外小时，将由付费服务用户使用，主要来自罗斯大楼和其他需要活细胞室的霍普金斯用户。

DESCRIPTION (provided by applicant): This application requests a Zeiss 510/META laser scanning confocal microscope with environmental chamber to replace our 11 year old Zeiss 410 microscope, which is part of the Imaging Core (Core Director, O. Kovbasnjuk, Ph.D., who is PI of this application) of an NIH NIDDK Core Development Center (R24, DK064388, "Hopkins Basic Science Digestive Diseases Core Center", PI, M. Donowitz, M.D.). The Zeiss 510/META microscope will allow us to provide state-of-the-art fluorescence imaging capabilities for 5 major interacting investigators, who are supported by the NIH NIDDK Core Development Center and who are closely interacting. There are also 6 minor users supported by the same Core Center who have their laboratories in the Ross Research Building, which houses the GI Core Center and the Zeiss confocal. In addition to being part of an NIH funded Core Center, the 510/META will be a resource for other JHUSOM scientists needing an environmental chamber on a confocal and will be available on a fee-for-service basis to scientists in a major Hopkins research facility (Ross Building). The 410 no longer satisfies our users' needs which can be met by the 510/META. This is due to 1) Deterioration of the Zeiss 410 PMT's and pin holes plus its outmoded software and tenuous computer support so the entire system will not be functional once the current computer fails. 2) Lack of spectral scanning capability needed for FRET applications. 3) Need for extensive use of FRET and FRAP with improved precision on 510 due to more precise pixel alignment than 410. The features of the Zeiss 510/META which are necessary to accomplish the diverse biomedical projects and which do not exist on the Zeiss 410 model to be replaced include: multitracking technology for speed and precise pixel localization; Zeiss LSM510 and Axiovision software for rapid signal processing and analysis; and a live cell chamber with temperature and CO2 controls needed for prolonged live cell imaging. The JHUSOM Imaging Core is not able to provide adequate time for the physiologic studies proposed in the R24 Center. This is preventing the proposed FRET studies and overnight constant temperature chamber studies. Importantly, no instruments available on a fee-for-service basis at JHUSOM are equipped with a live cell chamber. The Zeiss 510/META will be installed in the NIH NIDDK Core Center Imaging Core Facility, a location that is central to most of the users. It is anticipated that 75% of the 40 hr week will be used by the 5 major plus 6 minor users, with the remaining 25%, representing ~10-15 additional hrs/week, will be taken up by fee-for-service users, mostly from the Ross Building plus other Hopkins users needing the live cell chamber.