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

点击翻译按钮获取中文摘要

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.