Robinson G, Dawson I
Histochem J. 1975 Jul;7(4):321-33. doi: 10.1007/BF01007017.
This paper describes and immunochemical localization of gastrin-containing endocrine cells in the human pyloric antrum using antibodies conjugated to horseradish peroxidase. With pre-fixed cryostat sections, a distinct clear-cut staining of gastrin-containing cells can be obtained either by direct or indirect single stain procedures, but may cells containing endogenous peroxidase activity also stain. In order to abolish staining due to endogenous peroxidase, sections were pretreated with a number of inhibitors prior to incubation in immune sera, but the inhibitors used appeared to interfere with the antigenicity of the gastrin molecule since subsequent immunochemical localization was impossible. The application of a double-staining technique, however, allowed us to distinguish easily between those cells which contained endogenous peroxidase and those on to which labelled antibody had been adsorbed. No labelled cells were found in post-fixed cryostat sections of fresh-frozen tissue. The technique is of value because preparations are permanent, a fluorescence microscope is not required, and the same technique can be adapted for use with the electron microscope.
本文描述了使用与辣根过氧化物酶偶联的抗体对人幽门窦中含胃泌素的内分泌细胞进行免疫化学定位。对于预固定的冰冻切片,通过直接或间接单染程序可获得含胃泌素细胞明显清晰的染色,但含内源性过氧化物酶活性的细胞也可能染色。为了消除内源性过氧化物酶引起的染色,在免疫血清孵育前用多种抑制剂对切片进行预处理,但所用抑制剂似乎干扰了胃泌素分子的抗原性,因为随后无法进行免疫化学定位。然而,双重染色技术的应用使我们能够轻松区分含内源性过氧化物酶的细胞和吸附了标记抗体的细胞。在新鲜冷冻组织的后固定冰冻切片中未发现标记细胞。该技术具有价值,因为制备物是永久性的,不需要荧光显微镜,并且相同技术可适用于电子显微镜。