Confocal microscopy replacement

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This proposal requests funding for accessories that will add confocal capabilities to a recently purchased microscope. The goal is to allow the study in yeast cells of RNA decay in real time. The co-applicant laboratories and their collaborators are studying a broad range of questions dealing with the trafficking and stability of RNA and RNA protein complexes in both budding and fission yeasts. The overall aim is to understand how RNA stability and localization influence gene expression and regulate important cellular functions like the cell division cycle, genome stability, ribosome biogenesis and responses to changes in growth conditions. All members of the group use yeast as a model system and study dynamic RNA expression. They all require a special microscopy setup that allows strong magnification and rapid scanning to capture RNA expression and decay in real time. Until few months ago, we were using a first generation Olympus confocal microscope that provides proper magnification but lacks the speed necessary to capture very dynamic molecules. Unfortunately, after 10 years and due to the extensive use the mechanical motor of this microscope stopped working. The company no longer manufactures spare parts. We do not have any alternative source to complete ongoing experiments and several projects funded by NSERC are on hold. A purchase of new system is well over the limit of NSERC grant budgets and there is no other open competition at this moment. Therefore, we propose here to buy just the confocal module and attach it to an existing epifluorescence microscope. In this way, we will be able to continue our research and enhance existing multiuser equipments.