UCSF High Throughput Microscope

描述（由申请人提供）：此共享仪器拨款申请是用于最先进的高通量显微镜，将促进加州大学旧金山分校15名确定的研究人员的研究。两个已建立的核心基金的资源将为该文书的运行和维护提供大量支持。该计划将允许更多的美国国立卫生研究院在加州大学旧金山分校支持的研究项目使用该设备（2004年有833项研究资助）。制造商已同意将维修合同的价格降低20%。这将降低操作成本，促进高通量研究影响生物反应和药物作用的生化途径的复杂性和串扰。美国国立卫生研究院已将高通量研究和能力确定为其路线图的关键要素。揭示、分析和理解生物复杂性依赖于进行具有大量变量的高度多元研究的能力。信号的终末期标记，甚至是信号级联本身的直接相互作用、构象和亚细胞分配，都很容易使用荧光显微技术来测量。例如，申请人研究人员常规使用抗体染色来跟踪用激素、生长因子和分子或药物制剂处理的细胞、干细胞或器官培养物中多种荧光标记因子的数量和位置。在一些研究中，荧光团标记的蛋白质或细胞的数量和位置随时间在活细胞或器官培养中被跟踪。我们还通过测量荧光蛋白之间转移的能量量，对不同荧光蛋白标记的蛋白质的相互作用和构象进行了常规表征。我们的研究现在要求我们跟踪多种（甚至大量）生长因子、激素、药理学制剂和分子过表达/敲除的细胞效应。然而，传统显微镜严重限制了人工收集和分析图像所需的时间，从而限制了重要的研究。所选择的高通量显微镜系统Cellomics ArrayScan VTI具有集成板处理、机器人化合物输送、数据分析和数据库管理功能，将使我们的研究人员能够在几个小时内完成以前需要几个月的工作。在三维器官或细胞多孔培养中，孵育室将允许活细胞成像，载脂蛋白体附着将允许可选的特定光学切片收集。简而言之，拟议设备的安装将极大地提高我们的研究人员快速深入了解各种人类疾病的途径和干预措施的能力。

DESCRIPTION (provided by applicant): This Shared Instrument Grant application is for a state-of-the-art High Throughput Microscope that will facilitate the studies by 15 defined investigators at the University of California San Francisco. Resources from two established Core Facilities will provide substantial support for the operation and maintenance of this Instrument. This plan will permit access to this equipment by an even larger number of NIH-supported research projects at UCSF (833 Research Grants in 2004). The manufacturer has agreed to lower the price of maintenance contracts by 20%. This will reduce operating costs to facilitate high throughput studies of the complexities and cross-talk in biochemical pathways that affect biologic response and drug action. The NIH has identified high throughput studies and capabilities as a key element of its Roadmap. Uncovering, analyzing and understanding biologic complexities depend upon the ability to conduct highly multiplexed studies with large numbers of variables. End-stage markers of signaling and even the direct interactions, conformation and subcellular partitioning of components of the signaling cascade itself, are readily measured using fluorescent microscopic techniques. For example, the applicant investigators routinely use antibody staining to track the amounts and locations of multiple fluorescent-tagged factors in cell, stem cell or organ cultures treated with hormones, growth factors and molecular or pharmacologic agents. For some studies, the amounts and locations of fluorophore-marked proteins or cells are tracked with time in live cell or organ cultures. We also routinely characterize the interactions and conformations of proteins flagged with different fluorescent proteins through measurements of the amounts of energy transferred between the fluorescent proteins. Our studies now demand that we follow the cellular effects of multiple (and even large libraries of) growth factors, hormones, pharmacologic agents and molecular over- expressions/knock-downs. However, conventional microscopy severely constrains the amount of time required to manually collect and analyze images, thereby limiting important studies. The High Throughput Microscopy system selected, the Cellomics ArrayScan VTI with integrated plate handling, robotic compound delivery, data analysis and database managing capabilities, will allow our investigators to accomplish in hours what previously had required months of effort. An incubation chamber will permit live cell imaging, and an Apotome attachment will permit the optional collection of specific optical sections, in three dimensional organ or cell multi-well cultures. In short, the installation of the proposed equipment would dramatically increase the ability of our investigators to make rapid and deep inroads into the understanding of pathways and interventions for a wide variety of human diseases.