A High-performance Shared Resource Super-resolution Spinning Disk Microscope

A. Project Summary We request funds to purchase an Evident Scientific (Olympus) SpinSR, super-resolution spinning disk confocal microscope. This spinning disk microscope will replace and update our now obsolete technology, and will also provide a gentle, high-speed super-resolution method for live and fixed samples that our current structured illumination microscopy platform (end-of-life January 2024) cannot provide. The SpinSR will be housed within the Porter Biosciences Light Microscopy Core Facility (LMCF), located at the heart of the University of Colorado Boulder main campus. Here we identify 12 Major Users and 3 Minor Users, from 4 departments, who contribute to this proposal and will use the SpinSR. This User Group represents 18 active NIH awards (9 RO1, 5 R35, 1 DP, 1 R21 and 1 PO1,) and 1 HHMI award. In 2015, we assembled a spinning disk microscope around a repurposed Yokogawa CSU-10A Nipkow disk unit, which was built in August 2002. In 2017 we purchased a demonstration unit Yokogawa CV1000 spinning disk microscope that was built in 2011 and uses a Yokogawa CSU-X1 disk unit. We’ve maintained these microscopes, and they have been essential for our Users, having supported at least 39 peer-reviewed publications and numerous grants, doctoral and undergraduate honors theses, most from NIH-funded laboratories. As these microscopes are becoming dysfunctional (detailed in Rationale section) and are severely limited by today’s standards, our Users are increasingly unable to conduct their experiments, and they cannot even pursue certain experiments due to limitations of technology. Compared to our current spinning disk confocal microscopes, the SpinSR is more efficient, with a larger FOV and faster performance, uses 6 lasers, has 2 cameras for simultaneous 2-wavelength imaging, performs gentle, high frame-rate, real-time super-resolution, has expanded range high-speed optical sectioning, multipoint photo-bleaching/photo-stimulation and superior optical performance. The science represented in our User Group is diverse, ranging from mechanisms of proteasome quality control to – pair bonding in adolescents to – neural tube biology – illustrating our need for a flexible and robustly configured spinning disk microscope. Our current spinning disk and super-resolution capabilities cannot be serviced or updated, the microscopes and computers depend on unsupported software and operating systems and cannot fulfill our current and future needs, severely limiting our users. The SpinSR will address all the shortcomings of our current systems and enable our Users to extend their research by offering new capabilities and technologies, helping them to be more productive and make new discoveries, leading to high-impact papers and successful NIH grant proposals.