Acquisition of an automated live cell imaging platform.

点击翻译按钮获取中文摘要

The NSERC-funded research programs of principal investigators from the Health Sciences, Neuroscience, Biochemistry and Biology departments of Carleton University (CU) have been hindered by the absence of a live-cell imaging platform on campus. The original and highly innovative molecular biology, immunology and microbiology research programs of at least 6 internationally-recognized scientists rely heavily on cell imaging to study diverse biological questions and are compromised without availability of an automated, high-content live-cell microscope system. A critical issue is that our current microscopes limit users to end-point analyses of cell phenotypes. These end-point analyses require setting up parallel experiments for acquisition of multiple time points, provide limited coverage for each experiment, and are resource, time, and labor intensive. Most critically, end-point analyses do not fully reflect the dynamic nature of biological processes and are prone to human errors and therefore reduced experimental reproducibility. Execution of experiments with an automated platform will dramatically increase the number of experimental conditions (and cell lines) that can be analyzed in a single run and increase reproducibility. The ability to collect and analyze cell growth in real-time and simultaneously identify the outcomes of the treatments on the various cells (e.g. ROS, cytotoxicity, apoptosis, senescence) using commercially-available fluorescent reagents will maximize data collection, reduce cost, and greatly accelerate the research of all co-applicants. In addition, the real-time continuous imaging of cell viability, proliferation, growth, caspase activation and other phenotypes will also ensure that we never miss a phenotype, which is a constant concern when performing end-point assays. In this application we request funds for the acquisition of an IncuCyte S3 live-cell imaging platform for several research groups at Carleton University. The requested system is an affordable and easy-to-use automated microscope platform with the capacity to significantly advance fundamental molecular and cellular biology research at Carleton University. The S3 platform allows for straightforward identification of phenotypic changes by recording and tracking cell growth, cellular morphology, migration and invasion, or activation of specific pathways as a function of time in response to specific stimuli or inhibitors. The IncuCyte S3 platform will thus fill an important technological gap at Carleton University by providing real-time kinetic analysis of cell behavior over long time course experiments. This infrastructure will also allow us to foster new research partnerships within our institution and with external collaborators locally, nationally, and internationally. Consequently, this will greatly improve the funding competitiveness of our research programs at the national and international level, enhance our research capabilities and training of HQPs.