FLOW CYTOMETRY IN CANCER BIOLOGY AND IMMUNOLOGY

肿瘤研究越来越多地应用流式细胞术技术来研究肿瘤细胞和正常细胞的物理和功能特征。随着流式细胞术在基于荧光和光散射分离细胞方面的能力的发展，癌症研究的临床和基础科学家都使用这种仪器来分析和分类不同的细胞群。膜、细胞质和细胞核抗原表达、各种光散射特性、DNA含量和细胞周期分析、细胞凋亡检测以及细胞内生化变化如钙通量和pH变化的测量等参数均属于当前流式细胞术技术的范畴。最近，荧光蛋白的发展，可以在不同启动子的控制下表达，扩大了流式细胞术的能力，并为分子生物学家提供了一个强大的工具来分类细胞亚群。这里概述的每一种方法都代表了本提案中概述的癌症免疫学和分子生物学项目，以支持流式细胞仪设备，该设备支持马基癌症中心的癌症中心成员。这是一个完善的设施，对癌症中心研究项目的发展至关重要。该设施目前包含FACSCalibur和FACStar Plus，并将在1999年春季升级为MoFlo或Vantage SE。虽然我们期望继续得到癌症中心和大学的支持，但NIH对该设施的支持将使我们能够保持提高教师使用率和设施人员稳定性的费率。

Cancer research has increasingly applied the techology of flow cytometry to study the physical and functional characteristics of neoplastic and normal cells. As flow cytometry has developed its capabilities to separate cells based on fluorescence and light scatter, both clinical and basic scientists in cancer research have used this instrumentation to analyze and sort different populations of cells. Such parameters as membrane, cytoplasmic and nuclear antigen expression, various light scattering properties, DNA content and cell cycle analysis, the detection of apoptosis, as well as the measurement of intracellular biochemical changes such as calcium flux and changes in pH are within the realm of current flow cytometry technology. More recently, the development of fluorescent proteins which can be expressed under control of different promoters has expanded the capabilities of flow cytometry and provided a powerful tool for molecular biologists to sort subpopulations of cells. Each of the approaches outlined here is represented in the cancer immunology and molecular biology projects outlined in this proposal to support a Flow Cytometry Facility which supports the Cancer Center members at the Markey Cancer Center. This is a well established facility which has been essential to the development of Cancer Center research programs. The facility currently contains a FACSCalibur and FACStar Plus and will be upgraded in the Spring, 1999 with either a MoFlo or a Vantage SE. While we anticipate continued support from the Cancer Center and the University, NIH support for the facility will enable us to maintain rates that promote faculty usage and stability of the facility personnel.