Jáksó P, Pajor L
Pécsi Orvostudományi Egyetem, Patológiai Intézet.
Orv Hetil. 1998 Oct 18;139(42):2509-13.
The application of the CD45 vs. side scatter gating in the flow cytometric immunophenotyping of malignant haematological disorders was investigated. In the course of the conventional double labeling the light scatter based gating is the method of choice, which allows the immunophenotypic characterization of the subpopulations of interest. Using this gating strategy one can separate the lymphocytes, monocytes, neutrophil granulocytes, eosinophils and platelets as distinct cell populations in normal blood samples. The blastic cells present in different malignant hematological disorders might, however, represent an overlapping population with the lymphocytes, NK-cells, monocytes, and nucleated erytroid elements on the FSC vs. SSC histogram, making it difficult or impossible to separate them from the residual reactive cells. This will impart false mixed immunophenotypic property to the blastic cells. The authors found definitive advantages of CD45 vs. SSC gating in contrast to the FSC vs. SSC analysis due to the reliable separation of lymphoblasts, myeloblasts, nucleated erytroblasts as well as the debris from the reactive cell populations. The most plausible application of this method is shown in the course of three-color labeling, applying CD45 as a constant marker. Furthermore, the authors demonstrated that the CD45 vs. SSC gating strategy will result in considerable increase in resolution of blastic population also in the course of the most conventional dual-color labeling method.
研究了CD45与侧向散射门控在恶性血液系统疾病流式细胞术免疫表型分析中的应用。在传统的双重标记过程中,基于光散射的门控是首选方法,它可以对感兴趣的亚群进行免疫表型特征分析。使用这种门控策略,可以在正常血液样本中将淋巴细胞、单核细胞、中性粒细胞、嗜酸性粒细胞和血小板分离为不同的细胞群体。然而,不同恶性血液系统疾病中存在的原始细胞在FSC与SSC直方图上可能与淋巴细胞、NK细胞、单核细胞和有核红细胞成分重叠,使得难以或无法将它们与残留的反应性细胞分离。这将赋予原始细胞错误的混合免疫表型特征。与FSC与SSC分析相比,作者发现CD45与SSC门控具有明显优势,因为它能可靠地将淋巴母细胞、髓母细胞、有核红细胞以及碎片与反应性细胞群体分离。该方法最合理的应用体现在三色标记过程中,将CD45作为恒定标记物。此外,作者证明,在最传统的双色标记方法过程中,CD45与SSC门控策略也将显著提高原始细胞群体的分辨率。