Beavis A J, Pennline K J
Department of Immunology, Schering Plough Research Institute, Kenilworth, New Jersey 07033.
Cytometry. 1994 Apr 1;15(4):371-6. doi: 10.1002/cyto.990150413.
Multicolour immunofluorescence and flow cytometry were used for simultaneous measurement of five-cell surface antigens on murine spleen cells. We have been able to quantitate T-cells, T-cell subsets, B cells, and expression of the activation marker I-Ad from a single sample using four directly conjugated monoclonal antibodies LYT2-APC, L3T4-PE, B220-RED613, I-Ad-FITC and one indirect step THY1.2-biotin/streptavidin-Cascade Blue. Three excitation wavelengths were used (488 nm, 647 nm, and U.V. 351-364 nm) for fluorescence measurements. The combination of fluorochromes used provided good resolution such that all five fluorescence signals were spectrally resolved. The percentage of cells positive for expression of a specific cell surface marker were almost identical for single-colour samples and the five-colour analysis, differing by only 0.3-1.5 percentage points.
采用多色免疫荧光和流式细胞术同时检测小鼠脾细胞上的五种细胞表面抗原。我们能够使用四种直接偶联的单克隆抗体LYT2-APC、L3T4-PE、B220-RED613、I-Ad-FITC和一个间接步骤THY1.2-生物素/链霉亲和素-级联蓝，从单个样本中定量T细胞、T细胞亚群、B细胞以及活化标志物I-Ad的表达。使用三种激发波长（488nm、647nm和紫外351 - 364nm）进行荧光测量。所使用的荧光染料组合提供了良好的分辨率，使得所有五个荧光信号在光谱上得以分辨。特定细胞表面标志物表达阳性的细胞百分比在单色样本和五色分析中几乎相同，差异仅为0.3 - 1.5个百分点。
