Zeiss 510/META Confocal Microscope Plus Live Cell Chamber

描述(由申请人提供)：本申请请求蔡司510/META激光扫描共聚焦显微镜与环境室，以取代我们的11年历史的蔡司410显微镜，这是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.)成像核心的一部分。蔡司510/元显微镜将使我们能够为5名主要的互动研究人员提供最先进的荧光成像能力，他们得到NIH NIDDK核心开发中心的支持，正在密切互动。还有6个小用户由同一个核心中心支持，他们的实验室设在罗斯研究大楼，这是GI核心中心和蔡司共焦的所在地。除了是NIH资助的核心中心的一部分，510/META还将为其他需要共焦环境舱的JHUSOM科学家提供资源，并将以收费的方式向霍普金斯大学一个主要研究设施(罗斯大楼)的科学家提供服务。410不再满足我们用户的需求，而510/META可以满足这些需求。这是由于1)蔡司410PMT和针孔的退化，加上其过时的软件和薄弱的计算机支持，因此一旦当前的计算机出现故障，整个系统将无法运行。2)缺少FRET应用所需的光谱扫描能力。3)需要在510上广泛使用FRET和FRAP以提高精度，因为比410更精确的像素对齐。蔡司510/META是完成各种生物医学项目所必需的，而将被取代的蔡司410型号所不具备的功能包括：用于快速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LSM510和Axiovision软件；以及具有延长活细胞成像所需的温度和二氧化碳控制的活细胞室。JHUSOM成像核心不能为R24中心建议的生理学研究提供足够的时间。这阻碍了拟议的FRET研究和过夜恒温箱研究。重要的是，JHUSOM在收费服务的基础上提供的仪器都没有配备带电电池室。蔡司510/META将安装在NIH NIDDK核心中心成像核心设施中，该位置是大多数用户的中心。预计每周40小时中的75%将由5个主要用户和6个次要用户使用，其余25%，即每周增加约10-15个小时，将由收费服务用户占用，主要来自罗斯大楼和其他需要带电牢房的霍普金斯大学用户。

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.