Grant: Research Tools and Instruments

State-of-the-art microscopy provides our team with the ability to visualize cellular details and dynamic biological processes occurring in real time, allowing us to explore how cells and tissues behave using innovative tools and approaches. In this application, we are requesting upgrades to our Nikon Ti2-E CSU-XI spinning disk confocal microscope, a system that allows us to perform live imaging of cells and developing model organisms, like zebrafish, with minimal phototoxicity to living specimens. We have used our current equipment to explore questions related to how the heart develops and functions (Collins, principal applicant, ECR), describe the mechanisms that modulate neuronal excitability (Campanucci lab, co-applicant), and uncover the function of new epithelial cell types in the lung (Ianowski, co-applicant). Our research programs rely on access to live microscopy to investigate fundamentals aspects of cell biology and physiology. 36 HQP from 12 labs that use the spinning disk confocal microscope for an average of 50 hours per week. We are, however, facing multiple challenges to get full benefit from this instrument. We lack an external analysis computer, so users must book time on the microscope computer, delaying access for image acquisition for all labs. Our system lacks critical components to perform certain types of imaging, including a live cell incubator and optics for better 3D resolution of tissues. This impacts the applicant, co-applicants', and collaborators ability to perform our NSERC-focused research programs. To address these limitations, we are requesting funds for a live cell imaging upgrade, an analysis workstation and software upgrade, differential interference contrast (DIC) components, and a 40x silicone objective. These components will improve user access, allow us to image sensitive physiological processes like calcium dynamics in cardiac cells, and improve axial resolution and penetration depth for imaging deeper into tissues like organoids or the developing zebrafish heart. Our equipment is well-used and provides an excellent training opportunity for HQP in our labs. The Collins lab's spinning disk confocal is the only one in Saskatchewan and serves an unmet need for live imaging at USask. HQP use this equipment independently after receiving hands-on training sessions, and are supported during experiment planning, image acquisition, data analysis, and troubleshooting by experienced users in my lab and Nikon support staff. This unique training opportunity helps HQP to develop competitive expertise in advanced microscopy and image analysis that positions them with highly marketable skills. Investments to improve our existing infrastructure will allow us to continue to support HQP in our labs and accelerated our innovative research programs.