Zanyk M J, Banerjee D, McFarlane D L
Department of Pathology, University of Western Ontario, London, Canada.
Anal Cell Pathol. 1990 Jul;2(4):241-51.
A modified method of analysis by flow cytometry has been adapted to the NK/target cell system to monitor conjugate formation within a mixed population. The two-parameter assay is based on the use of two colour fluorescent markers in which an indirect FITC-conjugated NKH1A antibody (green fluorescence) is used to detect NK-specific effector cells and an intracellular strain, hydroethidine (red fluorescence) is used to detect target cells. The two cell populations and their conjugates are discernible by the presence of both green and red fluorescence, using a single laser beam tuned to 488 nm. In comparison, a manual count conjugate assay was applied using a hemacytometer and fluorescence microscope. Of the two methods, the automated technique using flow cytometry provides a lower but more precise representation of conjugate formation, due to the avoidance of technical/observer error that is common to manual count assays. The cytometric method has proven to be reproducible and superior in consistency to the manual count assay and can be adapted to almost any effector/target system under investigation. In comparison with previous methods using dual laser, the single laser system provides advantages of cost and time efficiency due to setup simplicity, as well as improved availability.
一种改进的流式细胞术分析方法已应用于NK/靶细胞系统，以监测混合群体中的共轭物形成。双参数检测基于使用两种颜色的荧光标记，其中间接异硫氰酸荧光素（FITC）偶联的NKH1A抗体（绿色荧光）用于检测NK特异性效应细胞，而细胞内染料羟基乙啶（红色荧光）用于检测靶细胞。使用调谐到488nm的单束激光，通过绿色和红色荧光的存在可区分这两种细胞群体及其共轭物。相比之下，使用血细胞计数器和荧光显微镜进行手动计数共轭物检测。在这两种方法中，由于避免了手动计数检测中常见的技术/观察者误差，使用流式细胞术的自动化技术提供了较低但更精确的共轭物形成表征。流式细胞术方法已被证明具有可重复性，并且在一致性方面优于手动计数检测，并且几乎可以适用于任何正在研究的效应器/靶细胞系统。与以前使用双激光的方法相比，单激光系统由于设置简单，在成本和时间效率方面具有优势，并且可用性更高。
