Spinning Disk Confocal and Single Molecule Localization Microscope

项目摘要/摘要： 13名NIH资助的调查人员正在寻求共享仪器拨款计划的支持，以获得 和或蜻蜓转盘共聚焦显微镜，具有单分子定位能力。这种多式联运 显微镜将为Bio-One的用户带来前所未有的速度、分辨率、灵敏度和大视场-- 特拉华大学成像中心(UD)。旋转圆盘共聚焦显微镜是当今最先进的 与点扫描相比，减少光漂白和光毒性的活细胞显微镜方法 共聚焦显微镜。此外，三维单分子定位显微镜是一种尖端的超分辨率 与此相关的成像技术在生物医学研究中引起了广泛的兴趣。这两个都不是 特拉华州的研究社区目前可以使用这些技术，因此蜻蜓将填补 一次需要两个缺口。目前的申请针对的是来自10个主要用户和8个次要用户的重点研究项目 对所需显微镜有迫切需求的用户。由于其灵活性和多模式，没有 怀疑这一仪器在其整个生命周期内将继续满足各种研究项目的需要。 蜻蜓将影响我们主要用户的广泛研究领域。这包括卡普兰博士，他 将解决细胞细胞器如何移动和释放信号的基本问题；Elliott博士和Wang博士， 世界卫生组织将检查肌肉骨骼疾病方面的胶原纤维和骨骼机械传感； 世卫组织将研究红细胞和眼晶状体中的细胞骨架。其他主要用户包括格雷霍恩博士， 谁将测量发育中的肺的动态过程；贾博士，他检查组织工程替代品 对于组织修复和再生；Grimes和Neunuebel博士将研究细菌细胞壁的生物合成和 病原菌在宿主细胞中的存活和复制；Vlachos博士将量化流体动力学， 塔尼斯博士将寻求确定细胞外小泡生物发生的潜在机制。 蜻蜓将被安置在一座新的最先进的研究大楼里，里面的成像套间特别 专为高端光学显微镜设计。它将由特拉华州生物技术研究所(DBI)的S生物- 成像中心，这是一个研究中心，为UD和UD的150多个研究小组提供成像需求 更远一点。DBI和UD在提供非常强大的机构支持方面有着悠久的历史，并拥有 承诺的空间、人员、服务合同和行政支持。一项为期5年的财务计划已经完成 提供这些服务是为了确保长期可持续性。生物成像中心在提供以下服务方面有着良好的记录 出色且持续的用户支持和资源管理。四名工作人员，他们有超过75年的历史 结合荧光显微镜的经验，将为蜻蜓的用户提供帮助 实验设计、样品制备、数据采集和分析。

Project Summary / Abstract: 13 NIH-funded investigators are seeking support under the Shared Instrumentation Grant Program to obtain an Andor Dragonfly spinning disk confocal microscope with single molecule localization capability. This multimodal microscope will bring unprecedented speed, resolution, sensitivity and large field of view to the users of the Bio- Imaging Center at the University of Delaware (UD). Spinning disk confocal microscopy is today’s state-of-the-art method for live-cell microscopy due to its reduced photobleaching and phototoxicity, compared to point-scanning confocal microscopy. Furthermore, 3D single molecule localization microscopy is a cutting edge super-resolution imaging technique with that has been attracting widespread interest in biomedical research. Neither of these technologies are currently available to the research community in Delaware, and the Dragonfly is thus set to fill two needed gaps at once. The current application targets key research projects from 10 major users and 8 minor users who have a critical need for the requested microscope. Thanks to its flexibility & multimodality, there is no doubt that this instrument will keep meeting the needs of a wide range of research projects throughout its lifetime. The Dragonfly will impact a broad range of research fields from our major users. This includes Dr. Caplan, who will address fundamental questions on how cellular organelles move and release signals; Drs. Elliott and Wang, who will examine collagen fibrils and bone mechanosensing in respect to musculoskeletal disorders; Dr. Fowler who will study the cytoskeleton in red blood cells and ocular lenses. Other major users include Dr. Gleghorn, who will measure dynamic processes in the developing lung; Dr. Jia, who examine engineered tissue substitutes for tissue repair and regeneration; Drs. Grimes and Neunuebel will study bacterial cell wall biosynthesis and pathogenic bacteria survival and replication in host cells, respectively; Dr. Vlachos will quantify fluid dynamics, and Dr. Tanis will seek to determine the mechanisms underlying the biogenesis of extracellular vesicles. The Dragonfly will be placed in a new state-of-the-art research building, in which imaging suites were specifically designed for high-end light microscopes. It will be managed by the Delaware Biotechnology Institute (DBI)’s Bio- Imaging Center, which is a research hub that serves the imaging needs of over 150 research groups at UD and beyond. DBI and UD have a long established history of providing very strong institutional support, and have committed space, personnel, service contract, and administrative support. A 5-year financial plan has been provided to ensure long-term sustainability. The Bio-Imaging Center has a strong track record for providing excellent and sustained user support and resource management. Four staff members, who have over 75 years of combined experience in fluorescence microscopy, will provide assistance to the Dragonfly’s users in experimental design, sample preparation, data acquisition, and analysis.