Enhanced Capability of Spinning Disk Confocal Microscope

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Understanding cellular processes requires the observation of dynamic changes and molecular interactions at micro- and nanometric length scales. The development of imaging methodologies enabling these observations at high resolution and in real time, therefore, revolutionized the biological and biomedical sciences. However, unlike the observation of fixed specimens, imaging live cells came with challenges arising from the damaging effect of the imaging process proper on cell viability and from the necessity to control the cells' physiological behavior. Spinning disk confocal (SDC) microscopy is a state-of-the art fluorescence imaging technique that is gentle and rapid. Compared to conventional epi-fluorescence or point scanning confocal laser microscopy, SDC microscopy reduces the damaging effect of phototoxicity and captures even weak fluorescent signals at high speeds. SDC microscopy is therefore an indispensible tool for cell biologists and biomedical scientists. The community of life science researchers at McGill's Macdonald Campus makes intensive use of the SDC microscope housed in the campus' Multi-Scale Imaging Facility (MuSIF). The system was acquired by Dr. Geitmann in 2015 with an NSERC grant and has since been expanded and upgraded with additional imaging capabilities such as a Total Internal Reflection Fluorescence (TIRF) detector, and a high-performing sCMOS camera. In the present proposal, additional components are requested to 1) further enhance the versatility of the instrument, 2) ensure the instrument's longevity and 3) ascertain environmental sustainability. The requested items serve to provide constant environmental conditions for long-term time lapse imaging (environmental chamber), more flexibility in terms of fluorescent markers (additional laser line, emission filters, second camera), and high spatial resolution (100x objective). The thusly enhanced capabilities will accommodate a wider range of biological organisms and experimental designs. Importantly, the expected life time of the instrument will be significantly extended by replacing the 10-year old workstation, and by substituting the mercury-based light source by a new LED light-source. The suite of fluorescence-based imaging instruments at MuSIF is regularly booked by the members of nearly 20 research teams at Macdonald Campus from disciplines as diverse as plant science, parasitology, microbiology and food science. The upgrades to the SDC microscope are essential for the advancement of ongoing and planned research projects that benefit Canadians and contribute to the advancement of fundamental science. Among the research topics are plant morphogenesis, viral infection processes, toxicity of nanomaterials, inflammatory gastrointestinal diseases and intracellular bacteria. The upgraded instrument will be a crucial tool for HQP technical skills training and it supports the activities of numerous research programs funded through NSERC Discovery grants.