Wynford-Thomas D, Jasani B, Newman G R
Histochem J. 1986 Jul;18(7):387-96. doi: 10.1007/BF01675220.
A method is presented which allows correlative serial section analysis by light and electron microscopy of cell surface antigens in monolayer cultures. Sites of antigenicity are shown by deposition of diaminobenzidine after pre-embedding, immunoperoxidase immunocytochemistry. Osmication is replaced by the use of gold chloride which specifically enhances the electron density of diaminobenzidine. In addition gold chloride bound to diaminobenzidine survives embedding and provides the basis for a post-embedding photochemical amplification method. Immunostained cells are embedded in LR White by a rapid technique which preserves their structure and leaves them available for subsequent post-embedding immunocytochemistry. The method is illustrated by the demonstration of epidermal growth factor (EGF) receptors on the EGF receptor-rich human carcinoma cell line A431 using a well characterized monoclonal antibody raised against EGF receptor.
本文介绍了一种方法，可通过光镜和电镜对单层培养细胞表面抗原进行相关连续切片分析。在预包埋免疫过氧化物酶免疫细胞化学后，通过二氨基联苯胺的沉积显示抗原性位点。用氯化金替代锇酸，氯化金可特异性增强二氨基联苯胺的电子密度。此外，与二氨基联苯胺结合的氯化金在包埋后仍能保留，并为包埋后光化学放大方法提供基础。免疫染色的细胞通过一种快速技术包埋在LR White中，该技术可保留其结构并使其可用于后续的包埋后免疫细胞化学。使用针对表皮生长因子（EGF）受体的特征明确的单克隆抗体，在富含EGF受体的人癌细胞系A431上证明了EGF受体，从而对该方法进行了说明。
