Abstract We are requesting funds to purchase a Zeiss LSM 980 confocal fluorescence microscope with Airyscan 2. This instrument combines super-resolution, fast scanning, and high signal-to-noise imaging modes that will greatly expand the imaging capabilities at the University of Rochester’s High Content Imaging Core (HCIC). Access to the instrument will be made available to a user base that extends throughout the University of Rochester. Many research topics at the University of Rochester encounter questions that cannot be answered with conventional confocal microscopy due to the diffraction limit on resolution, the bleaching rates of scarce fluorophores, high phototoxicity, or a combination of these factors. Over the past 15 years, a variety of innovative super-resolution techniques have been developed that allow optical microscopy to explore details below the diffraction limit. However, techniques that boost resolution typically come with a cost in speed (e.g. single molecule localization microscopy (SMLM)) and/or phototoxicity (e.g. simulated emission depletion (STED) microscopy). These costs can be too high for many biological applications. Airyscan is a super-resolution technique that has become popular in recent years because it offers improved resolution (~2x increase) along with higher temporal resolution and lower phototoxicity relative to conventional laser scanning confocal microscopes. This technology would therefore provide enormous benefit to a wide range of ongoing projects at the University of Rochester. We have established an initial user group comprised of researchers from across the University of Rochester River Campus. These scientists span a diverse set of research interests that cover cutting edge areas of quantitative biology including cell differentiation, phase separated DNA condensates, functional genomics, the molecular basis of aging, and cell trafficking. The Zeiss LSM 980 with Airyscan 2 is an ideal resource to support these investigators and future researchers in advancing the goals of their NIH-funded research.

Project Narrative Both human disease and healthy human development are driven by intricate events occurring at the nanoscale within cells. The Zeiss LSM 980 with Airyscan 2 uses cutting-edge developments in fluorescent microscopy to let researchers at the University of Rochester peer inside the cell at smaller length scales, faster time scales, and for longer periods of time. This technology will have a transformative impact on ongoing NIH-funded research into how developing organisms form proper tissues, which molecular elements drive evolution, how pathogens cross from the bloodstream into tissues, how cells organize their DNA.