A platform for live-cell time-lapse imaging

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RTI funding will be used to assemble a state-of-the-art wide-field fluorescence microscopy platform for time-lapse imaging of live yeast cells. The platform will enable sophisticated analysis of protein localization, abundance, and turnover dynamics, as well as spatial and temporal analysis of protein-protein interactions. The platform will be unique in that it will be dedicated to long-term time-lapse imaging using microfluidic devices, a capability that is currently lacking in Toronto, where instrument time on shared microscopes or in shared facilities is either impossible to book due to time constraints, or is prohibitively expensive. The platform is critically required as increasingly sophisticated cell biological analyses encompass the temporal component and study individual cells rather than population phenomena. Research conducted by major users focuses on single-cell models, particularly budding yeast, to study diverse areas of biology including genome integrity and transmission, regulation of protein location and abundance, autophagy, quantitative genetics, and regulation of mRNA translation and localization. The demand for an imaging platform with the capability of performing time-lapse experiments for hours up to days, in controlled environments, is high among the major users, ensuring maximum utilization. We anticipate that the availability of the platform will enable increasingly sophisticated analyses by minor users in the Biochemistry and Molecular Genetics departments and in the Donnelly Centre. A wide range of applications are anticipated, particularly protein fluorescence-based and cell morphology-based phenotypic assays that incorporate temporal and environmental condition components.