Spinning Disk Confocal and Single Molecule Localization Microscope

项目摘要/摘要：

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.