A platform for live-cell time-lapse imaging

RTI资金将用于组装一个最先进的广域荧光显微镜平台，用于对活的酵母细胞进行延时成像。该平台将实现对蛋白质定位、丰度和周转动态的复杂分析，以及对蛋白质-蛋白质相互作用的空间和时间分析。该平台的独特之处在于，它将致力于使用微流控设备进行长期延时成像，这是多伦多目前缺乏的一种能力，在多伦多，共享显微镜或共享设施上的仪器时间要么由于时间限制无法预订，要么昂贵得令人望而却步。随着日益复杂的细胞生物学分析涵盖时间部分并研究单个细胞而不是群体现象，该平台是至关重要的。主要用户的研究主要集中在单细胞模型，特别是萌芽酵母，以研究生物学的不同领域，包括基因组完整性和传递、蛋白质位置和丰度的调节、自噬、数量遗传学以及信使核糖核酸的翻译和定位调节。主要用户对能够在受控环境中进行长达数小时甚至数天的延时实验的成像平台的需求很高，以确保最大限度地利用。我们预计，该平台的提供将使生物化学和分子遗传学部门以及唐纳利中心的小用户能够进行越来越复杂的分析。预计会有广泛的应用，特别是基于蛋白质荧光的和基于细胞形态的表型分析，它结合了时间和环境条件的成分。

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.