A live-cell analysis system for multiplex quantitation and fluorescent imaging

O‘Donoghue博士是化学生物学的CRC，他的NSERC资助的研究团队设计了不同类型的荧光记者，以量化活细胞中蛋白质合成和酶活性的错误。该实验室开发了新的荧光记者，可以实时、具有空间和时间分辨率地可视化氧化还原或激酶活性。利用活细胞记者的能力依赖于基础设施来支持细胞的高内容和高通量成像。这个应用程序需要紧急资金，用于最先进的细胞成像和分析系统Cytation C10。在西方大学，没有C10或类似的仪器具有C10的所有功能。Cytation C10将放置在我们的BioCore设施中，供部门和西部大学更大的研究社区使用，仪器将在那里进行专业管理，用户费用将包括实际操作的HQP培训和维护费用。该设施服务于500个HQP的用户群，所需的仪器对至少150个HQP和40个WESTERN研究小组来说是迫切需要的。BioCore工厂已经可持续运行了25年，包括专职的管理人员和技术人员，以确保设备的正确使用、培训和维护。由于我们的设施为学生提供实践培训，C10将直接供HQP使用，并为HQP提供一个宝贵的机会，让他们接受关于现有最先进的活细胞成像系统的培训。在同一个实验中，我们在获取高含量成像和高通量荧光测量的能力和能力方面面临着一个关键的瓶颈，这将通过收购C10来解决。包括我们的申请者、合作者和BioCore用户团队，Cytation 10将推进我们由NSERC支持的哺乳动物细胞生物学和生物化学不同领域的研究目标，包括蛋白质合成、细胞信号和RNA代谢的研究。Cytation C10能够对细胞过程和表型进行量化，通过在标准孵化器中全天候自动收集和分析图像来解决随时间推移的问题。重要的是，C10实现了西方大学所没有的几种先进方法和方法：i)通过将活细胞高通量多模式平板阅读器与高内容成像仪相结合，ii)通过提供灵活的荧光读取和成像以允许更广泛的分析和荧光记者，以及iii)通过定位我们的设施来升级到用于高分辨率3D成像和单细胞图像的共聚焦显微镜。总而言之，Cytation C10将消除研究的一个关键瓶颈，并极大地扩展我们的实验能力，以承担涉及生物分子和活细胞中酶活性可视化的具有挑战性的项目。数以百计的研究人员和HQP将从这项先进技术中受益。

Dr. O'Donoghue is a CRC in Chemical Biology and his NSERC-funded research team engineers fluorescent reporters of diverse types to quantify errors in protein synthesis and enzyme activity in living cells. The lab develops new fluorescent reporters to visualize redox or kinase activity in real time and with spatial and temporal resolution. The ability to utilize live-cell reporters relies on infrastructure to support high-content and high-throughput imaging of cells. This application requests urgent funding for a state-of-the-art cellular imaging and analysis system, the Cytation C10. There is no C10 or similar instrument with all the capabilities of the C10 at Western University. The Cytation C10 will be situated in our BioCore facility for use in the department and the larger research community at Western, where the instrument will be professionally managed and user fees will cover hands-on HQP training and maintenance costs. The facility serves a user base of >500 HQP, and the requested instrument is of immediate need for at least 150 HQP and 40 research groups at Western. The BioCore facility has run sustainably for 25-years and includes dedicated managers and technicians to ensure proper use, training, and maintenance of equipment. Because our facility provides hands-on training to students, the C10 will be used directly by HQP and provides an invaluable opportunity for HQP to receive training on the most advanced live-cell imaging system available. We face a critical bottleneck in capacity and ability to acquire high-content imaging and high-throughput fluorescent measurement in the same experiment that will be resolved by acquisition of the C10. Inclusive of our team of applicants, collaborators, and BioCore users, the Cytation 10 will advance our NSERC-supported research objectives in diverse areas of mammalian cell biology and biochemistry, including studies on protein synthesis, cellular signaling, and RNA metabolism. The Cytation C10 enables quantitation of cellular processes and phenotypes, resolved over time by automatically gathering and analyzing images around the clock within a standard incubator. Critically, the C10 enables several advanced methods and approaches that are not available at Western University: i) by combining a live-cell high-throughput multi-mode plate reader with a high-content imager, ii) by providing flexibility in fluorescent reading and imaging to allow a broader range of assays and fluorescent reporters as well as enable engineering new fluorescent protein reporters, and iii) by positioning our facility to upgrade to confocal microscopy for 3D imaging and single-cell images at high-resolution. In summary, the Cytation C10 will remove a critical bottleneck to research and vastly expand our experimental capabilities to undertake challenging projects involving visualization of biomolecules and enzyme activities in living cells. Hundreds of researchers and HQPs will benefit from this advanced technology.