Nakane P K, Hartman A L
Histochem J. 1980 Jul;12(4):435-47. doi: 10.1007/BF01011959.
A method for the localization of intracellular antigens with a scanning electron microscope using peroxidase-labelling antibodies is described. A search for a hydrogen donor which may be deposited at the sites of antigen by enzymatic action and emit secondary electrons or generate backscatter electrons was made. It was found that when 4-chloro-1-naphthol was used, the peroxidase deposited reaction product which resulted in a strong secondary electron emission at the site of antigen. With this method, the presence of luteinizing hormone in secretion granules and other cytoplasmic structures of gonadotropic cells was demonstrated. The level of detection of intracellular antigens with this method is not as high as that detectable with light microscopical examination of the same specimens, that is, more reaction product at the site of antigen is required to be detectable with scanning electron microscopy than with light microscopy. In spite of the lack of high sensitivity, the intracellular antigens may be localized with the method described.
描述了一种使用过氧化物酶标记抗体通过扫描电子显微镜定位细胞内抗原的方法。研究了寻找一种氢供体，它可通过酶促作用沉积在抗原部位，发射二次电子或产生背散射电子。发现当使用4-氯-1-萘酚时，过氧化物酶沉积反应产物在抗原部位产生强烈的二次电子发射。用这种方法，证明了促性腺细胞的分泌颗粒和其他细胞质结构中存在促黄体生成素。用这种方法检测细胞内抗原的水平不如对相同标本进行光学显微镜检查时可检测到的水平高，也就是说，与光学显微镜相比，扫描电子显微镜需要在抗原部位有更多的反应产物才能检测到。尽管缺乏高灵敏度，但仍可使用所述方法定位细胞内抗原。
