Confocal microscopy replacement

本提案要求为最近购买的显微镜增加共聚焦能力的配件提供资金。目标是允许在酵母细胞中实时研究RNA衰变。共同申请实验室及其合作者正在研究有关出芽和裂变酵母中RNA和RNA蛋白复合物的运输和稳定性的广泛问题。总体目标是了解RNA稳定性和定位如何影响基因表达和调节重要的细胞功能，如细胞分裂周期、基因组稳定性、核糖体生物发生和对生长条件变化的反应。该小组所有成员都以酵母为模型系统，研究动态RNA表达。它们都需要一种特殊的显微镜装置，允许强大的放大和快速扫描来实时捕捉RNA的表达和衰变。直到几个月前，我们还在使用第一代奥林巴斯共聚焦显微镜，它提供了适当的放大倍率，但速度不够快，无法捕捉非常动态的分子。不幸的是，10年后，由于广泛使用，这台显微镜的机械马达停止工作。这家公司不再生产零配件了。我们没有任何替代来源来完成正在进行的实验，NSERC资助的几个项目被搁置了。购买新系统远远超过NSERC拨款预算的限制，目前没有其他公开竞争。因此，我们建议在这里只购买共聚焦模块，并将其连接到现有的荧光显微镜。这样，我们将能够继续我们的研究和提高现有的多用户设备。

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.