Parker D J, Haslam P L
Cell Biology Unit, Cardiothoracic Institute, London, U.K.
J Immunol Methods. 1988 May 25;110(1):37-45. doi: 10.1016/0022-1759(88)90080-4.
A method is described for the removal of contaminating erythrocytes from immunofluorescence labelled lymphocyte preparations and allowing accurate gating on the lymphocyte population during flow cytometric analysis. The technique involves an improved erythrolytic fixative combined with the DNA stain propidium iodide permitting non-nucleated cells (i.e., red blood cells) to be gated out during analysis. The erythrolytic fixative was shown not to be detrimental to the major T cell markers detected using indirect immunofluorescence. This method will improve the analysis of blood lymphocytes and their subsets in mononuclear cell preparations and more particularly in samples of body fluids and tissue washings or extracts which may contain variable and often large numbers of erythrocytes.
本文描述了一种从免疫荧光标记的淋巴细胞制剂中去除污染红细胞的方法,该方法能在流式细胞术分析过程中对淋巴细胞群体进行准确设门。该技术涉及一种改良的红细胞裂解固定剂,结合DNA染料碘化丙啶,使无核细胞(即红细胞)在分析过程中被设门排除。结果表明,这种红细胞裂解固定剂对使用间接免疫荧光检测的主要T细胞标志物没有损害。该方法将改善对单核细胞制剂中血液淋巴细胞及其亚群的分析,特别是对可能含有数量可变且往往大量红细胞的体液、组织冲洗液或提取物样本的分析。