INSTRUMENTATION FOR FUNCTIONAL CYTOMETRY

我的研究计划是关注新的细胞计数技术的癌症研究和医学的发展。同一肿瘤内的细胞在分子组成和功能特性上不同。因此，研究和诊断癌症需要以单细胞分辨率分析细胞的分子组成和功能特性。用于单细胞分辨率定量分析的技术统称为细胞计数法。直到最近，还没有用于分析细胞的分子组成和功能特性的细胞计数技术。我们通过开创性的化学和功能性细胞分析技术缩小了这一技术差距。化学细胞计数法现在是一种成熟的方法，被许多研究实验室广泛使用。相反，功能性细胞计数是我们在过去3年中提出的一种新方法，有望彻底改变癌症研究和癌症医学。我们在几篇论文中描述的原理验证实验是通过使用基本设备以每天约30个细胞的速度进行的，这比功能性细胞计数法的实际应用所需的速度低100万倍。在这里，我们建议建立一个功能性细胞计数仪，每天可以分析数百万个细胞，从而促进这项新技术的进一步发展和实际应用。该仪器基于高容量成像仪和先进的图像处理软件。当收购时，该仪器将被用于其全部能力，因为支持的研究计划是非常好的资助（NSERC和CIHR赠款总额超过$700，000每年）和充分的工作人员（5研究生，3 PDF，和4研究助理）。我们将在许多重要项目中使用这种仪器，包括开发基于耐药性的癌症亚型的技术。后一项技术将促进个性化的癌症治疗，有可能为加拿大人的健康和福祉做出重大贡献。如果实验室不能在2015年获得拟议的仪器，我们的许多研究方向将受到阻碍，有些将完全停滞。这反过来又会导致研究生完成学位要求的时间大大推迟。

My research program is concerned with the development of novel Cytometry techniques for cancer research and medicine. The cells within the same tumor differ in their molecular composition and functional properties. Studying and diagnosing cancer, therefore, require analysis of molecular composition and functional properties of cells with single-cell resolution. Techniques used for quantitative analyses with single-cells resolution are collectively called Cytometry. Up until recently, there were no Cytometry techniques for analyses of molecular composition and functional properties of cells. We have closed this technological gap by pioneering Chemical and Functional Cytometries. Chemical Cytometry is now a well established approach widely used by many research laboratories. Functional Cytometry is, in contrast, a new approach laid down by us in the last 3 years, which promises to revolutionize cancer research and cancer medicine. Our proof-of-principle experiments, described in several papers, were performed by using rudimentary equipment with a speed of ~30 cell per day, which is 1 million times less than required for practical applications of Functional Cytometry. Here we propose to build an instrument for Functional Cytometry that will allow the analysis of millions of cells per day and will, thus, facilitate further development and practical use of this novel technology. The instrument is based on a high-content imager and advanced image processing software. When acquired, the instrument will be used to its full capacity as the supported research program is very well funded (NSERC and CIHR grants totaling over $700,000 per year) and fully staffed (5 graduate students, 3 PDFs, and 4 research associates). We will use this instrument for many important projects, including the development of technology for drug-resistance-based cancer subtyping. The latter technology will facilitate personalized cancer treatment with the potential to significantly contribute to the health and wellbeing of Canadians. If the laboratory is not able to obtain the proposed instrument in 2015, many directions of our research will be impeded and some will be completely stalled. This, in turn, will cause significant delays in completion of degree requirements by graduate students.