Super-resolution spinning disk confocal microscope for Dartmouth Life Sciences

用于达特茅斯生命科学的超分辨率转盘共焦显微镜

• 批准号: 10427997
• 负责人: Yasmath Ahmed
• 金额: $ 60万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427997
• 关键词: Animals; Biological Sciences; Cell Culture Techniques; Cells; Color; Communities; Confocal Microscopy; Disease; Ensure; Equipment; Funding; Future; Goals; Grant; Image; Kidney Diseases; Laboratories; Lasers; Malignant Neoplasms; Mammalian Cell; Microscope; Microscopy; Minor; Mycoses; Noise; Optics; Photons; Phototoxicity; Physiology; Recording of previous events; Research; Research Personnel; Resolution; Sampling; Services; Signal Transduction; Speed; System; Technology; Tissue Sample; Training; United States National Institutes of Health; Yeasts; autism spectrum disorder; college; developmental disease; experience; fluorescence imaging; fly; imaging capabilities; innovation; instrument; light microscopy; medical schools; temporal measurement

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.

项目摘要 来自达特茅斯学院五个部门和盖塞尔医学院的十二名NIH资助的调查员 请求资金购买尼康的横川CSU-W1 Sora旋转磁盘共聚焦显微镜（Nikon） Sora）可以实现荧光活细胞和组织的快速，超分辨率的多通道成像 样品。请求的系统以每秒200帧的速度集成了同时进行的两色成像，x- Y分辨率为120 nm，由XY Galvo高速扫描仪和四个启用了动态光刺激 激光器（405 nm，488 nm，561 nm和640 nm）。该系统适用于广泛的现场和固定样品 包括整个动物后生系统（Fly，蠕虫），哺乳动物细胞培养和酵母在内的应用。八 主要用户和四名未成年人的支持得到18款NIH赠款的支持，以进行针对的创新研究 阐明正常生理和疾病状态的机制，包括癌症，自闭症， 发育障碍，退化性肾脏疾病和真菌感染。达特茅斯的现有系统 不能满足对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（Nikon Sora）是满足主要/次要用户的这些未满足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）为高速，超级 - 现有设备目前无法实现的现场和固定样品的分辨率成像；和2）到 长期降低光毒性和光损伤，而无需牺牲分辨率和信噪比 敏感或弱表达的活样品的成像。通过在旋转中添加光学光子重新分配 磁盘技术并使用反卷积来优化分辨率，尼康Sora系统提供了独特的 超分辨率成像与高速采集的组合。请求的共聚焦显微镜系统 因此，将是达特茅斯的一种独一无二的工具，并彻底改变了成像能力 达特茅斯生活科学社区。显微镜将被集中放置并维护 Life Sciences光学显微镜设施，它在支持生命科学研究的悠久历史上很长，并且是 由50多个实验室使用，这些实验室在很大程度上得到了支持，从而确保了所有人的广泛访问 达特茅斯的研究人员和对NIH资助的研究的强烈影响。达特茅斯学院和盖斯尔学校 医学已为这次购买提供了100,000美元的机构支持，以及三个服务的服务 具有丰富经验的成像专家在提供成像支持和培训方面，这将确保 要求的Sora显微镜系统仍然受到支持，多年来，达特茅斯研究人员可用于 来。