Super-resolution spinning disk confocal microscope for Dartmouth Life Sciences

项目摘要 来自达特茅斯学院和盖泽尔医学院五个系的12名NIH资助的研究人员 申请资金从尼康（Nikon）购买Yokogawa CSU-W1索拉旋转圆盘共聚焦显微镜 索拉），能够对荧光活细胞和组织进行快速、超分辨率、多通道成像 样品所要求的系统集成了同步双色成像，每秒高达200帧，X- Y分辨率为120 nm，通过XY振镜高速点扫描仪和四个 激光（405 nm、488 nm、561 nm和640 nm）。该系统适用于各种活的和固定的样品 应用包括整个动物后生动物系统（蝇、蠕虫）、哺乳动物细胞培养和酵母。八 主要用户和四个次要用户得到18个NIH赠款的支持，以进行创新研究， 阐明了正常生理和疾病状态，包括癌症，自闭症， 发育障碍、退行性肾病和真菌感染。达特茅斯的现有系统 不能满足这些未来扩展所需的高空间和时间分辨率的需求。 项目尼康索拉是满足主要/次要用户这些未满足的成像需求的最佳解决方案 并被选为实现两个密切相关的技术目标：1）提供一个解决方案，高速，超， 目前现有设备无法实现的活样本和固定样本的分辨率成像;以及2） 在不牺牲长期分辨率和信噪比的情况下降低光毒性和光损伤 敏感或弱表达的活样品的成像。通过将光子重新分配添加到自旋中， Nikon索拉系统采用了独特的磁盘技术，并使用反卷积来优化分辨率， 超分辨率成像与高速采集的结合。所要求的共聚焦显微镜系统 因此，这将是达特茅斯独一无二的仪器，并彻底改变成像能力， 达特茅斯生命科学社区。显微镜将安装在位于中心的 生命科学光学显微镜设施，在支持生命科学研究方面有着悠久的历史， 由50多个实验室使用，这些实验室主要由NIH支持，从而确保所有人都能广泛使用 达特茅斯的研究人员和强大的影响NIH资助的研究。达特茅斯学院和盖泽尔学校 医学部已承诺为此次购买提供10万美元的机构支持，以及三个 在提供成像支持和培训方面具有丰富经验的成像专家，这将确保 要求的索拉显微镜系统仍然支持和提供给达特茅斯研究人员多年， 来了

PROJECT ABSTRACT Twelve NIH-funded investigators from five departments at Dartmouth College and the Geisel School of Medicine request funds to purchase a Yokogawa CSU-W1 SoRa spinning disk confocal microscope from Nikon (Nikon SoRa) that would enable rapid, super-resolution, multi-channel imaging of fluorescent live cell and tissue samples. The requested system integrates simultaneous two-color imaging at up to 200 frames per second, X- Y resolution to 120 nm, dynamic photostimulation enabled by an XY galvo high-speed point scanner and four lasers (405 nm, 488 nm, 561 nm, and 640 nm). This system is suitable for a wide range of live and fixed sample applications including whole animal metazoan systems (fly, worm), mammalian cell culture, and yeast. The eight Major Users and four Minor Users are supported by 18 NIH grants to conduct innovative research aimed at elucidating the mechanisms that underlie normal physiology and disease states, including cancer, autism, developmental disorders, degenerative kidney disease and fungal infection. Existing systems at Dartmouth do not fulfill the needs for both high spatial and temporal resolution necessary for the future expansion of these projects. The Nikon SoRa is an optimal solution to fulfill these unmet imaging needs of the Major/Minor Users and was selected to achieve two closely related technical goals: 1) to provide a solution for high-speed, super- resolution imaging of live and fixed samples that is currently unattainable with existing equipment; and 2) to reduce phototoxicity and photodamage without sacrificing resolution and signal-to-noise ratio for the long-term imaging of sensitive or weakly expressing live samples. By adding optical photon reassignment to the spinning disk technology and using deconvolution to optimize resolution, the Nikon SoRa system provides a unique combination of super-resolution imaging with high-speed acquisition. The requested confocal microscopy system would therefore be a one-of-a-kind instrument at Dartmouth and revolutionize imaging capability for the Dartmouth life sciences community. The microscope will be housed in and maintained by the centrally-located Life Sciences Light Microscopy Facility, which has a long history of supporting life sciences research and is utilized by over 50 laboratories that are largely NIH-supported, thereby ensuring widespread access for all Dartmouth investigators and strong impact on NIH-funded research. Dartmouth College and the Geisel School of Medicine have committed $100,000 in institutional support for this purchase, as well as the services of three imaging experts with extensive experience in providing imaging support and training, which will ensure that the requested SoRa microscopy system remains supported and available for Dartmouth researchers for years to come.