Super-resolution spinning disk confocal microscope for Dartmouth Life Sciences

项目摘要

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.