Wang B L, Larsson L I
Histochemistry. 1985;83(1):47-56. doi: 10.1007/BF00495299.
Available techniques for light and electron microscopical double immunocytochemical staining are all associated with certain problems. We have developed a novel multiple staining procedure, which allows use of antibodies of differing specificities, raised in the same species (e.g. rabbit). Its essential features include 1) saturation of antigenic epitopes on the first layer primary antiserum by second (fluorophor- or gold-) labelled anti-IgG antibodies and 2) denaturation of free anti-IgG binding sites by formaldehyde vapour treatment. Various combinations of gastrin, somatostatin, glucagon, ACTH, growth hormone and enkephalin/endorphin antibodies have been tested at the light and electron microscopical level and have been found to give highly reproducible double- and triple-staining results. The technique has also been evaluated by use of cytochemical paper models. The method is simple and very useful for multiple staining of a wide variety of antigens.
现有的光镜和电镜双重免疫细胞化学染色技术都存在一定问题。我们开发了一种新型多重染色程序，该程序允许使用在同一物种（如兔）中产生的具有不同特异性的抗体。其主要特点包括：1）用第二种（荧光或金）标记的抗IgG抗体饱和第一层抗血清上的抗原表位；2）通过甲醛蒸汽处理使游离抗IgG结合位点变性。已在光镜和电镜水平上测试了胃泌素、生长抑素、胰高血糖素、促肾上腺皮质激素、生长激素和脑啡肽/内啡肽抗体的各种组合，发现可产生高度可重复的双重和三重染色结果。该技术也已通过细胞化学纸模型进行评估。该方法简单，对于多种抗原的多重染色非常有用。
