Dynamic Imaging: high resolution, long-term approaches to molecular mechanisms

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Watching cells as they react to conditions, divide, and die gives us important information that can't be measured in other ways. We request a microscope that takes detailed pictures over long periods of time while keeping cells alive; our current facilities cannot support this work because they are booked solidly and are not suitable. This equipment is important because tracking individual cells during environmental stress- such as starvation, temperature change, or drug treatments- gives information that influences Canadian agriculture, livestock and the environment.******Our labs study the basic molecular pathways and cellular structures that influence three important areas. First, Sarah Sabatinos asks how DNA is protected or mutated by environmental stress and drug treatments. Second, Roberto Botelho and Gagan Gupta observe organelle formation, maturation and roles in cell health. Third, Costin Antonescu tests how cells sense and respond to starvation or nutrition, and the role that this plays in cell growth and division. Together, we will use a variety of proteins that are fused to glowing fluorescent proteins. These fluorescent fusions are monitored using the live cell microscope to watch patterns over time. For example, tagged chromosomes can detect if DNA is altered by temperature and drug treatments. Lysosomes, the cell's eating machines, can be watched as they form, fuse and change. A cell's cilia are hair-like structures, that we watch form and grow. In these assays, we will use fluorescent tags to screen unknown populations and discover new gene interactions and protein functions that can be tested in later years.******The microscope that we request is optimized for these experiments; it takes high-resolution images, and has low-light levels and an environmental conditioning chamber which helps to keep cells alive during the movie-making process. Additionally, it is able to photograph and track multiple different conditions over long periods of time, a “high-content” process.******The recent growth of both our labs, our Department and our graduate program has increased the number of undergraduate, graduate and postdoc students who would use this microscope. This requested equipment will be used solidly and constantly with the work we describe. We are severely limited in our ability to perform these kinds of cutting edge research because we lack this requested equipment. Our 2 current microscopes are fully booked and cannot be used for long movies. They are also not appropriate for these experiments, nor can they acquire high-content data.******Our students will benefit tremendously from this equipment- it will be used to complete their theses, write papers, collaborate between our labs and internationally. Our students gain valuable skills in microscopy in this work, and help us to create new ways to analyze data. This training gives our HQP an advantage that will translate into jobs, and new research directions.