NSF/FDA SIR: Isolation of Cancer Stem Cells by Flow Cytometric Techniques

该研究的主要目的是采用先进的基于流式细胞术的仪器来开发分离不同细胞亚群的方法和方案。利用荧光检测对细胞特征进行深入分析，在单个细胞上同时区分17种或更多特征，并采用多种参数对这些群体进行分类，同时保持细胞的活力，这将促进流式细胞仪在生物医学研究中的应用。癌症祖细胞的分离和鉴定直到最近才成为可能，因为技术已经发展到由几个激光器发射的特定波长激发的多种荧光染料和独特的探测器可以同时根据多种标记对细胞进行分类。这需要稳定的激光线，仔细的补偿控制，确定的流体和敏感的光学和探测器。要验证的假设是，基于可识别的特征，可以识别慢性淋巴细胞白血病（CLL）中最早的癌症祖细胞。

ABSTRACT #1238375RavecheThe major objective of the proposed research is to employ advanced flow-cytometry-based instrumentation to develop methods and protocols for isolating distinct subpopulations of cells. This in-depth analysis of cell characterizations using fluorescence detection to distinguish 17 or more characteristics simultaneously on individual cells and employing multiple parameters to sort these populations while maintaining the viability of the cells will advance the employment of flow cytometers in biomedical research. The isolation and identification of the cancer progenitor cell is only recently feasible because the technology has evolved such that multiple fluorochromes excited by specific wavelengths emitted from several lasers and unique detectors can sort cells based on multiple markers simultaneously. This requires stable laser lines, careful compensation controls, defined fluidics and sensitive optics and detectors. The hypothesis to be tested is that based on identifiable characteristics, the earliest cancer progenitor cell in chronic lymphocytic leukemia (CLL) can be identified.