De Waele M, De Mey J, Moeremans M, De Brabander M, Van Camp B
J Histochem Cytochem. 1983 Mar;31(3):376-81. doi: 10.1177/31.3.6186731.
Colloidal gold was used as a marker for the light microscopic detection of lymphocyte cell surface antigens with monoclonal antibodies. Suspensions of peripheral blood leukocytes were first incubated with monoclonal mouse antibodies and then with colloidal gold-labeled goat anti-mouse antibodies. The cells were fixed and cytocentrifuge preparations or smears were made. Granulocytes and monocytes were then labeled by the cytochemical staining of their endogenous peroxidase activity. Lymphocytes reacting with the monoclonal antibody had numerous dark granules around the surface membrane. With electron microscopy, these granules appeared as patches of gold particles. This immunogold staining method proved to be a reliable tool for the enumeration of T-lymphocyte subpopulations in peripheral blood. The results were almost identical to those obtained with immunofluorescence microscopy. The procedure can also be applied on small volumes of capillary blood. This constitutes a good microtechnique for the determination of lymphocyte subsets in children.
胶体金用作标记物，通过单克隆抗体对淋巴细胞表面抗原进行光学显微镜检测。首先将外周血白细胞悬液与小鼠单克隆抗体孵育，然后与胶体金标记的山羊抗小鼠抗体孵育。细胞固定后，制作细胞离心涂片或涂片。然后通过粒细胞和单核细胞内源性过氧化物酶活性的细胞化学染色对其进行标记。与单克隆抗体反应的淋巴细胞在表面膜周围有许多深色颗粒。通过电子显微镜观察，这些颗粒表现为金颗粒斑块。这种免疫金染色方法被证明是对外周血中T淋巴细胞亚群进行计数的可靠工具。结果与免疫荧光显微镜获得的结果几乎相同。该方法也可应用于少量毛细血管血。这构成了一种用于测定儿童淋巴细胞亚群的良好显微技术。
