Sato S, Adachi A, Sasaki Y, Ghazizadeh M
Central institute for Electron Microscopic Researches, Nippon Medical School, Tokyo, Japan.
J Microsc. 2008 Jan;229(Pt 1):17-20. doi: 10.1111/j.1365-2818.2007.01881.x.
In conventional transmission electron microscopy, uranyl acetate staining is used to enhance the cellular components. However, uranyl acetate is considered a radioactive material that is very toxic if ingested or inhaled and subject to restrictions in many countries. In an attempt to introduce a substitute for uranyl acetate, we evaluated oolong tea extract (OTE) for staining of ultrathin sections. Tissue sections from normal rat liver representing an ideal model organ were processed according to a routine electron microscopic fixation and embedding procedure. Serial ultrathin sections were cut and processed with either routine double electron staining or 0.2% OTE staining for 30-40 min at room temperature followed by lead citrate staining (OTE staining method). Transmission electron microscopy observations revealed that all sub-cellular structures in hepatocytes were clearly visible with OTE staining and the quality of staining was highly compatible with those of routine double staining methods. It is suggested that OTE could be used as a non-radioactive and hazard-free substitute for uranyl acetate in transmission electron microscopy staining.
在传统透射电子显微镜术中,醋酸铀酰染色用于增强细胞成分。然而,醋酸铀酰被认为是一种放射性物质,摄入或吸入时毒性很大,在许多国家受到限制。为了尝试引入醋酸铀酰的替代品,我们评估了乌龙茶提取物(OTE)用于超薄切片染色的效果。将来自正常大鼠肝脏的组织切片作为理想的模型器官,按照常规电子显微镜固定和包埋程序进行处理。切取连续超薄切片,分别采用常规双电子染色或在室温下用0.2% OTE染色30 - 40分钟,随后进行柠檬酸铅染色(OTE染色法)。透射电子显微镜观察显示,用OTE染色时肝细胞中的所有亚细胞结构都清晰可见,且染色质量与常规双染色方法高度兼容。这表明OTE可作为透射电子显微镜染色中醋酸铀酰的无放射性且无危害的替代品。