Mansour I, Jarraya M A, Gane P, Roquin H, Rouger P, Doinel C
Institut National de Transfusion Sanguine, CHP-INTS, Paris, France.
Cytometry. 1990;11(5):636-41. doi: 10.1002/cyto.990110512.
Many laboratories do not have access to a flow cytometer allowing three-color immunofluorescence analysis through the use of multiple light sources. In view of the usefulness of such analyses in the dissection of cell parameters, we describe an approach permitting the study of three labels by using one light source and the two-color immunofluorescence assay. It is useful for the enumeration of cell subpopulations positive for one label and negative for two or more others as well as for qualitative analysis concerning the expression of these labels. This approach is simple and rapid; it does not require additional material and technical steps other than that used in the two-color immunofluorescence assay. Briefly, it consists of the use of a label coupled to a dye (PE or FITC or instance) and two different labels coupled to the other dye. An argon ion laser, operating at 488 nm and 60 mW, excites both fluorescein and phycoerythrin conjugated antibodies. We provided a general example, using three hypothetical labels (X, Y, and Z), and four practical applications: CD3+CD4CD8- and CD8+CD16-CD3- peripheral blood lymphocytes, CD2+CD16-CD3- and CD56+CD16-CD3- peripheral blood, and decidual infiltrating lymphocytes.
许多实验室无法使用通过多个光源进行三色免疫荧光分析的流式细胞仪。鉴于此类分析在剖析细胞参数方面的实用性，我们描述了一种通过使用单一光源和双色免疫荧光测定法来研究三种标记物的方法。它对于计数一种标记物呈阳性而另外两种或更多种呈阴性的细胞亚群以及对这些标记物表达的定性分析很有用。这种方法简单快速；除了双色免疫荧光测定法中使用的材料和技术步骤外，不需要额外的材料和技术步骤。简而言之，它包括使用与一种染料（例如PE或FITC）偶联的一种标记物以及与另一种染料偶联的两种不同标记物。一台波长为488nm、功率为60mW的氩离子激光器可激发荧光素和藻红蛋白偶联抗体。我们提供了一个使用三种假设标记物（X、Y和Z）的一般示例以及四个实际应用：CD3 + CD4CD8 - 和CD8 + CD16 - CD3 - 外周血淋巴细胞、CD2 + CD16 - CD3 - 和CD56 + CD16 - CD3 - 外周血以及蜕膜浸润淋巴细胞。
