Zeiss 510/META Confocal Microscope Plus Live Cell Chamber

描述（由申请人提供）：本申请要求Zeiss 510/Meta激光扫描共聚焦显微镜与环境室，以取代我们的11岁的Zeiss 410显微镜，这是成像核心的一部分（核心主任，O。Kovbasnjuk博士，他是本申请的PI）的NIH NIDDK核心开发中心（R24，DK 064388，“霍普金斯基础科学消化疾病核心中心”，PI，M. Donowitz，M.D.）。蔡司510/Meta显微镜将使我们能够为5个主要的相互作用的研究人员提供最先进的荧光成像能力，这些研究人员得到NIH NIDDK核心开发中心的支持，并且他们正在密切互动。还有6个小用户由同一个核心中心支持，他们的实验室位于罗斯研究大楼，该大楼设有GI核心中心和蔡司共聚焦。除了作为NIH资助的核心中心的一部分外，510/Meta还将成为需要共焦环境室的其他JHUSOM科学家的资源，并将以收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不再满足我们的用户需求，而510/Meta可以满足这些需求。这是由于1）蔡司410 PMT的退化和针孔加上其过时的软件和脆弱的计算机支持，使整个系统将无法正常工作，一旦目前的计算机故障。2)缺乏FRET应用所需的光谱扫描能力。3)需要广泛使用FRET和FRAP，由于比410更精确的像素对齐，510的精度有所提高。蔡司510/Meta的功能是完成各种生物医学项目所必需的，而蔡司410型号所不具备的功能包括：用于速度和精确像素定位的多跟踪技术;用于快速信号处理和分析的蔡司LSM 510和Axiovision软件;以及用于长时间活细胞成像所需的温度和CO2控制的活细胞室。JHUSOM Imaging Core无法为R24 Center拟定的生理学研究提供充足的时间。这阻碍了所提出的FRET研究和过夜恒温室研究。重要的是，JHUSOM的收费服务仪器都没有配备活细胞室。蔡司510/Meta将安装在NIH NIDDK核心中心成像核心设施中，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，剩余的25%（约10-15小时/周）将由按服务收费的用户使用，主要来自Ross Building以及需要活细胞室的其他霍普金斯用户。

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.