Maurer D, Felzmann T, Knapp W
Institute of Immunology, University of Vienna, Austria.
J Immunol Methods. 1990 Dec 31;135(1-2):43-7. doi: 10.1016/0022-1759(90)90254-s.
Here we describe a flow cytometry method which permits the collection of data regarding three staining parameters of a single cell from a suitable combination of dual parameter stainings using PE- and FITC-labelled mAbs and a single argon laser. The technique neither requires a third dye with non-overlapping emission spectra nor a second light source. In order to test the accuracy of this method, we compared data calculated using this triple parameter analysis with data obtained by double staining a presorted population homogeneously positive for one parameter. In experiments using either resting or in vitro activated T cells, the percentages obtained with both methods were identical (P greater than 0.05). Using this method we tested which T cell sub-subpopulation is responsible for the weak CD25 (IL-2R alpha) expression constantly associated with freshly isolated human T cells and concluded that it is predominantly expressed on the CD4+ CD45RO+ (CD4 positive memory) T cell subpopulation.
在此，我们描述了一种流式细胞术方法，该方法允许使用PE和FITC标记的单克隆抗体以及单个氩激光，从双参数染色的合适组合中收集关于单个细胞的三个染色参数的数据。该技术既不需要具有不重叠发射光谱的第三种染料，也不需要第二个光源。为了测试该方法的准确性，我们将使用这种三参数分析计算的数据与通过对预分选的一个参数均匀阳性的群体进行双重染色获得的数据进行了比较。在使用静息或体外活化T细胞的实验中，两种方法获得的百分比相同（P大于0.05）。使用这种方法，我们测试了哪个T细胞亚亚群负责与新鲜分离的人类T细胞持续相关的弱CD25（IL-2Rα）表达，并得出结论，它主要表达于CD4+ CD45RO+（CD4阳性记忆）T细胞亚群上。
