Sibon O C, Humbel B M, De Graaf A, Verkleij A J, Cremers F F
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Histochemistry. 1994 Mar;101(3):223-32. doi: 10.1007/BF00269548.
A high-resolution in situ hybridization method is described for localizing epidermal growth factor (EGF)-receptor transcripts in nuclei of A431 epidermoid carcinoma cells. The method is based upon the use of ultra-small gold particles in combination with silver enhancement. The RNA of the EGF-receptor was detected mainly around the nucleoli. After removal of the DNA using nucleases and high salt extraction, the RNA of the EGF-receptor appears to be associated with the nuclear matrix. The RNA of the EGF-receptor was observed in close contact with the SC-35 splicing protein, but no exact colocalization was observed. These results demonstrate that high resolution pre-embedding in situ hybridization in combination with immunocytochemistry, both using ultra-small gold as a detection method, provides a powerful tool to unravel the organization of nuclear processes.
描述了一种高分辨率原位杂交方法，用于在A431表皮样癌细胞的细胞核中定位表皮生长因子（EGF）受体转录本。该方法基于使用超小金颗粒结合银增强技术。EGF受体的RNA主要在核仁周围被检测到。使用核酸酶和高盐提取去除DNA后，EGF受体的RNA似乎与核基质相关。观察到EGF受体的RNA与SC-35剪接蛋白紧密接触，但未观察到精确的共定位。这些结果表明，结合免疫细胞化学的高分辨率预包埋原位杂交，两者均使用超小金作为检测方法，为揭示核过程的组织提供了一个强大的工具。
