University of Regina Cellular Impacts Facility Microscopes and Cell Culture

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Cell sorting and microscopy are well established methods for understanding the physiology of cellular systems, their response to xenobiotics and host-pathogen interactions, shared challenges of the applicants. The UR Cell Impacts Facility (CIF) in the Institute for Environmental Change and Society houses a platform for these studies. For example, the study of host-pathogen interactions first requires BL2 pathogens and host cells to be cultured, pathogen or host cells to be assessed/separated based on biomarkers using a high end flow cytometer/cell sorter, and finally host-pathogen interactions imaged (atomic force, confocal, fluorescence microscopies) of as a function of phenotypic and physiological changes. The proposed components and equipment would restore to full function this shared platform, essential to our NSERC-funded research programs, comprehensive HQP training, in-house and external collaborations and for retaining our national and international reputations. As the CIF equipment ages, a number of the instruments associated with this platform are in need of upgrading or replacement. The BL2 labs used to prepare samples for cell sorting/flow cytometry, mass spectral analysis and microscopy, are currently inoperable, necessitating an incubator-shaker for fungal cell culture and a microscope to assess human cell viability, confluence and quality. Validation of host cell status (cytometry) and enrichment of infected host cells (sorting) based on fluorescent biomarkers is not currently possible, requiring a new 488 nm main laser line to operate, and a 405 nm laser that excites dyes in the violet-blue. Host-pathogen interactions are then screened by fluorescence microscopy followed by confocal or correlative imaging to produce high-resolution 3D publication quality images of their interaction including the subcellular co-localization of labeled protein targets. The Zeiss 780 laser scanning confocal microscope currently has five continuous wave laser lines (458, 488, 514, 543, 594 nm) and a 63 x objective, precluding ultra-high resolution (100 x) imaging of dyes that excite in the violet-blue (405 nm). The research projects of our HQP are currently limited without a fully functioning platform, with certain experiments at a standstill and others having to be modified accordingly, ultimately slowing progress. The ability to use this platform to its full capacity is thus critically required to enable the co-applicants and their HQP to address ambitious research questions. Our applicant team and HQP emerge from a range of backgrounds that is reflective of current Canadian diversity, fueling our creativity, discovery and innovation. The equipment upgrades and replacements are indispensable tools that will allow this platform to operate at maximum capacity for many years, supporting research programs that impact both environmental and health sciences.