A Point-Scanning Confocal Microscope for the Biomedical Sciences

University Of Wisconsin-Madison, Madison WI

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Abstract

PROJECT SUMMARY In 2009, the University of Wisconsin (UW) was awarded an NIH Shared Instrumentation Grant to establish a confocal microscope in an imaging core facility. The core acquired a Nikon A1R confocal microscopy system and began immediately offering investigators from across the campus the opportunity to incorporate confocal imaging in their research. Here we request funds for a direct replacement of this 13-year-old instrument with a Nikon AX R confocal microscopy system equipped with both galvanometer and resonance scanners, four laser lines, spectral and high-sensitivity GaAsP detectors, autofocus and incubation systems for 4D time-lapse imaging, and Nikon Elements software. This instrument will be housed at the University of Wisconsin Optical Imaging Core (UWOIC) facility, where the A1R instrument has served 275 users from 90 laboratories in 31 departments across 5 schools. The UWOIC facility is well-supported institutionally by the UW School of Medicine and Public Health (UWSMPH) to provide access to a range of optical imaging modalities to all research groups on campus, with an emphasis on high speed, high-resolution microscopy. Importantly, UWSMPH has outfitted our core facility with a full tissue culture suite, with space for animal housing available on a nearby floor, to enable users from anywhere on campus to use instruments here. Funds committed to this application from the UWSMPH Dean’s office will provide salary support for the full-time UWOIC facility manager and greatly extend the service contract and therefore the lifetime of this new instrument. UWSMPH funded multiple software and hardware upgrades to the current A1R confocal to extend the warranty, but after 13 years Nikon considers this instrument obsolete and made the decision not to allow further extensions. Age, poor stability, and outdated and end of life optical and electrical components motivate the immediate acquisition of a new instrument. In selecting this replacement, microscopes from Leica Microsystems (Stellaris), Olympus (FV3000), Zeiss (LSM 980 Airyscan 2), and Nikon (AX R) were evaluated to determine which instrument would be the best suited for operation in our multi-user imaging facility. After comparing each of the systems mentioned and reviewing their reliability and service records, we concluded that the Nikon AX R offers the best balance of flexibility in scanning and detection capabilities, highest field of view, best software interface, and live-cell imaging performance needed to serve investigators with a wide range of research projects and experience. Each component of this system is well justified by the needs of more than 20 users that exemplify the depth and breadth of the research requiring confocal microscopy at our facility.