Spinning Disk Confocoal Microscope

描述(申请人提供)：克利夫兰临床勒纳研究所的成像核心和NIH资助的15名研究人员申请资金购买用于活细胞成像的旋转圆盘共聚焦显微镜系统-特别是与徕卡DMI6000倒置显微镜耦合的Perkin Elmer Ultraview Vox系统。LRI成像核心成立于15年前，旨在为克利夫兰临床研究社区提供尖端的数字成像设备和专业知识。该设施将容纳和管理所请求的仪器，使其在不被主要和次要用户使用时可供克利夫兰地区的科学界使用。LRI成像核心总监朱迪思·德拉兹巴博士拥有20多年的共焦经验，其中15人负责管理该设施。三名全职技术人员和一名研究助理协助她。Core有两个立式徕卡SP2共焦显微镜，目前有来自24个部门的122名用户大量使用。这些仪器仅限于对载玻片上的固定样品进行成像。目前在克利夫兰的任何地方都没有旋转圆盘共聚焦显微镜系统。所要求的仪器将为研究人员提供研究动态细胞过程所需的极大提高的时间分辨率。它还将提供进行荧光共振能量转移(FRET)测量、光漂白后荧光恢复(FRAP)测量和光激活的能力，从而大大推进他们由NIH资助的生物医学研究。主要使用者的项目包括：TIMP-3对血管生成的调节(Anand-Apte博士)，负责转移癌细胞中肌球蛋白II组装和定位的信号通路(Egelhoff博士)，肌动蛋白聚合在血管生成过程中调节细胞极化和迁移的作用(Fox博士)，质膜-肌动蛋白细胞骨架重构对B淋巴细胞激活、增殖和运动的调节(Gupta博士)，紧密连接和质膜之间的囊泡运输在调节血管内皮生长因子驱动的内皮细胞定向迁移中的作用(Horowitz博士)，G蛋白b2亚单位在转录调节过程中的作用，基因激活和染色质重塑(Karnik博士)，PAR-1在调节黑腹果蝇卵巢边缘细胞迁移过程中细胞骨架和黏附蛋白定位中的作用(McDonald博士)，以及Smoothened蛋白的靶向和转运(Smoothened Protein)，Hedgehog信号转导中的一个必不可少的激活剂(朱博士)。

DESCRIPTION (provided by applicant): The Imaging Core of the Cleveland Clinic Lerner Research Institute and a group of fifteen NIH- funded investigators request funds to purchase a spinning disk confocal microscope system for live-cell imaging - specifically the Perkin Elmer Ultraview Vox System coupled to a Leica DMI6000 inverted microscope. The LRI Imaging Core was established fifteen years ago to provide leading-edge digital imaging equipment and expertise for the Cleveland Clinic research community. This facility will house and manage the requested instrument, making it available to the Cleveland area scientific community when not being used by the major and minor users. Dr. Judith Drazba, LRI Imaging Core Director, has over twenty years' confocal experience, fifteen of them running this facility. Three full-time technicians and a Research Associate assist her. The Core has two upright Leica SP2 confocal microscopes, heavily used by 122 current users from 24 departments. These instruments are limited to imaging fixed samples on slides. There is currently no spinning disk confocal microscope system anywhere in Cleveland. The requested instrument will offer researchers greatly improved temporal resolution needed to study dynamic cellular processes. It will also provide the ability to do fluorescence resonance energy transfer (FRET) measurements, fluorescence recovery after photobleaching (FRAP) measurements and photoactivation, thereby substantially advancing their NIH-funded biomedical research. Projects of the major users involve regulation of angiogenesis by TIMP-3 (Dr. Anand-Apte), signaling pathways responsible for assembly and localization of myosin II in metastatic cancer cells (Dr. Egelhoff), role of actin polymerization regulating cell polarization and migration during angiogenesis (Dr. Fox), regulation of B lymphocyte activation, proliferation, and motility by plasma membrane-actin cytoskeletal remodeling (Dr. Gupta), role of vesicle trafficking between tight junctions and the plasma membrane in the regulation of VEGF-driven directional migration of endothelial cells (Dr. Horowitz), the role of the G protein b2 subunit in the process of transcription regulation, gene activation and chromatin remodeling (Dr. Karnik), the role of Par-1 in regulating the localization of cytoskeletal and adhesion proteins during ovarian border cell migration in Drosophila melanogaster (Dr. McDonald), and the targeting and trafficking of Smoothened protein, an essential activator in Hedgehog signaling transduction (Dr. Zhu).