High End Laser Scanning Confocal Microscope for the University of Chicago Integrated Light Microscopy Facility - Project Summary / Abstract The University of Chicago Integrated Light Microscopy Facility (ILMF) requests funds to purchase a high-end laser scanning confocal microscope. The ILMF currently serves 420 users in 80 labs from across the University. Sixty-seven of those labs use laser scanning confocal microscopy, and 78% of those labs have NIH funding. Usage hours have increased as the ILMF’s microscope capacity has decreased. Two of our confocal microscopes, both Leica SP5 models, are over 14 years old. Leica has designated them end-of-life, meaning they are no longer manufacturing parts for these systems and replacements are not guaranteed. We have already experienced failure of the 488nm Argon and 592nm depletion lasers on one, and failure of the Mai Tai multiphoton excitation laser on the other, with no possibility of replacing any of these components. We expect to decommission at least one SP5 within the next year, making users hesitant to start new projects on those systems. This has stressed our two newer laser scanning confocal systems (purchased with institutional funds in 2016 and 2020), pushing them to use levels averaging 91% of AUT, defined as 3640 hours per year. The system proposed here is the Evident (formerly Olympus) Fluoview 4000 (FV4000), released in 2024. The system will increase the capacity and functionality of laser scanning confocal microscopes in the ILMF, allowing users to collect high-quality data more readily. Several features of the FV4000 will be new to the ILMF, and satisfy a number of outstanding investigator needs. Features include: state-of-the-art, patented, fast signal processing silicon photomultiplier (SiPM, Evident SilVIRTM) detectors, to significantly improve signal-to-noise levels, enhancing detection of Golgi cisternae and other organelle sub-structures; four high magnification, long working distance silicone immersion objectives for detailed, multi-color, 3-dimentional imaging of organoids, thick tissues and tumor samples; and three near-infrared wavelength lasers for excitation of fluorophores beyond the current imaging spectrum, allowing for investigation of a larger number of molecules of interest in a single sample. The FV4000 will also feature full environmental control, allowing users to take advantage of faster imaging speeds to image live samples. This will make it possible to image longer sessions at higher frame rates with less photodamage, resulting in more robust and reliable data from live samples than currently possible. Finally, the FV4000 base is modular in design, allowing for field upgrades with Evident or third-party resources (e.g. a single molecule localization module) as users’ experimental needs grow. In summary, adding an Evident FV4000 laser scanning confocal microscope to the ILMF will make it possible for users to gather information from samples that are currently challenging but valuable research models.
