Spinning Disk Super Resolution Confocal Microscope for the Vanderbilt Brain Institute

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PROJECT SUMMARY / ABSTRACT Funds are requested to purchase a Nikon Spinning Disk Super Resolution (CSU-W1/SoRa) Confocal microscope for imaging projects focused around mental health research within the Vanderbilt Brain Institute (VBI), a multi-disciplinary research institute comprised of faculty at Vanderbilt University (VU) and Vanderbilt University Medical Center (VUMC). The requested instrument will be integrated into and managed by the Vanderbilt Cell Imaging Shared Resource (CISR), a microscopy core with a 20+ year history of successful microscopy management, training, and acquisition of new technology. This instrument capable of confocal imaging and super resolution imaging in live samples is ideal for 6 Major and 7 Minor Users within the VBI with projects surrounding synaptogenesis, synapse assembly and plasticity, subsynaptic signaling, and communication between cell and tissue populations. These molecular and cellular investigations will advance already funded NIMH and NIH projects totaling over $6.8M in FY22 in the areas of social interaction, impulsive aggression, learning and memory, Rett syndrome (RTT), ADHD, and more. The proposed microscope configuration is based on the needs of the VBI and Major Users for live imaging at cellular and subcellular resolution. The Nikon CSU-W1 SoRa confocal microscope includes: 1) CSU-W1 spinning disk plus Yokogawa SoRa super resolution spinning disk for super resolution near 120 nm in XY; 2) stage-top incubation for imaging live cells and organoids; 3) Z-Piezo stage for increased speed in Z for 3D volumes; 4) Hamamatsu Orca-Fusion sCMOS camera with 30-100 frames per second; 5) excitation lasers of 405, 488, 561, and 633 nm; and 6) objective lenses for live cell imaging at high resolution ranging from 10x, 20x, 40x, 60x, and 100x. This new system would be housed in existing CISR space in Medical Research Building III (MRBIII) where the VBI is based and the majority of the Major User labs are located. Vanderbilt University is providing financial support to promote the long-term operation of the instrument. If awarded, the Office of the Dean of Basic Sciences is committed to providing $75,000 for CISR staff salary, computer purchases and upgrades, and reduction in user fees over the first 5 years. Technical support and training of new users will be facilitated by the exceptional staff within the CISR, which has a greater than 20-year history of assisting over 300 labs on campus. Moreover, this equipment will become part of the Vanderbilt Nikon Center of Excellence (COE) that is housed within the CISR. This CISR/Nikon COE provides a dedicated local team of specialists to support the microscopy needs of investigators on campus. Collectively, the proposed confocal system, together with outstanding institutional and core facility support, will enable the long-term objective of supporting and advancing neuroscience and mental health research Vanderbilt.