Acquisition of an automated live cell imaging platform.

由于校园缺乏活细胞成像平台，来自卡尔顿大学(CU)健康科学、神经科学、生物化学和生物系的主要研究人员的NSERC资助的研究项目受到了阻碍。至少6名国际公认的科学家的原创和高度创新的分子生物学、免疫学和微生物学研究计划严重依赖细胞成像来研究各种生物学问题，如果没有自动化的、高含量的活细胞显微镜系统就会受到影响。一个关键的问题是，我们目前的显微镜限制了用户对细胞表型的终端分析。这些端点分析需要为获取多个时间点设置并行实验，为每个实验提供有限的覆盖范围，并且是资源、时间和劳动力密集型的。最关键的是，终点分析不能完全反映生物过程的动态性质，容易出现人为错误，因此降低了实验的重复性。使用自动化平台进行实验将极大地增加可在一次运行中分析的实验条件(和细胞系)的数量，并提高重复性。使用商用荧光试剂实时收集和分析细胞生长并同时识别对各种细胞的处理结果(例如，ROS、细胞毒性、细胞凋亡、衰老)的能力将最大限度地收集数据，降低成本，并极大地加速所有共同申请者的研究。此外，对细胞活力、增殖、生长、半胱氨酸天冬氨酸氨基转移酶激活等表型的实时连续成像也将确保我们永远不会错过表型，这是进行终点分析时持续关注的问题。在本申请中，我们申请资金为卡尔顿大学的几个研究小组购买IncuCyte S3活细胞成像平台。所要求的系统是一个负担得起且易于使用的自动化显微镜平台，能够显著促进卡尔顿大学的基础分子和细胞生物学研究。S3平台可以通过记录和跟踪细胞生长、细胞形态、迁移和入侵或作为特定刺激或抑制物响应时间函数的特定通路的激活来直接识别表型变化。因此，IncuCyte S3平台将填补卡尔顿大学的一项重要技术空白，提供对长时间过程实验中细胞行为的实时动力学分析。这一基础设施还将使我们能够在我们的机构内部以及与当地、国家和国际的外部合作者建立新的研究伙伴关系。因此，这将极大地提高我们在国家和国际层面的研究项目的资金竞争力，增强我们的研究能力和HQP的培训。

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.