McKeever P E, Garvin A J, Spicer S S
J Histochem Cytochem. 1976 Aug;24(8):948-55. doi: 10.1177/24.8.60441.
A method is described for ultrastructural localization of immune complex receptors on the surface of viable peritoneal exudate cells. The technique entails incubation with a soluble complex of horseradish peroxidase (HRP) and specific antibody to HRP at 4 degrees C followed by exposure to diaminobenzidine and processing for electron microscopy. The bound immune complexes were evident as focal deposits of HRP reaction product, adhering closely to the external surface of macrophages with an uninterrupted periodicity varying between 30 and 120 nm. Following incubation with an insoluble immune complex containing a higher proportion of antibody, receptor sites stained frequently, but large aggregates adhered to the cells. Rinsing cells after staining with soluble complexes partially displaced the bound immune complexes. Fixation prior to exposure to immune complexes largely eliminated the binding capacity of the immune complex receptors.
本文描述了一种用于在活腹膜渗出细胞表面超微结构定位免疫复合物受体的方法。该技术包括在4℃下与辣根过氧化物酶(HRP)和抗HRP特异性抗体的可溶性复合物孵育,然后暴露于二氨基联苯胺并进行电子显微镜处理。结合的免疫复合物表现为HRP反应产物的局灶性沉积,紧密附着于巨噬细胞外表面,不间断的周期性在30至120nm之间变化。用含有较高比例抗体的不溶性免疫复合物孵育后,受体位点频繁染色,但大的聚集体附着于细胞。用可溶性复合物染色后冲洗细胞可部分置换结合的免疫复合物。在暴露于免疫复合物之前进行固定可大大消除免疫复合物受体的结合能力。