钨酸钠作为稳定剂的新的高灵敏HRP-TMB法——Ⅱ、电镜研究

本文研究了各种不同锇化步骤对TMB-ST反应产物的影响。结果发现:TMB-ST法结合DAB-Co后处理步骤用于电镜束路追踪时,具有反应产物电子密度很高,呈特征性的细杆状,组织超微结构保存良好的特点,且灵敏度较高。而未后处理的TMB-ST产物锇化后大多数只具有中等电子密度。此外,通过对以硝普钠、钼酸铵、柠檬酸镍离子络合物、钨酸钠作为稳定剂的各种TMB法的比较,我们首次提出了稳定剂是影响TMB产物电子密度的最重要因素的观点。

A NEW HIGHLY SENSITIVE HRP-TMB METHOD USING SODIUM TUNGSTATE AS A STABILIZER: Ⅱ. ELECTRON MICROSCOPIC STUDY

Until now the developed TMB methods for electron microscopy (TMB-EM method) are all unable to result in maximum demonstration of transported HRP while maintaining good ultrastructure preservation. The present study showed that this problem can be solved when the TMB-ST method combined with DAB-Co procedure was adopted. In the electron microscopic retrograde and anterograde fiber tracing, the sensitivity was higher than other developed TMB-EM methods, meanwhile the ultrastructural preservation was adequate. The reaction products of the TMB-ST-DAB-Co procedure under electron microscopy presented a characteristic small rod-shaped structure which was extremely electron dense and was often aggregated in clumps. The results also show ed that stabilizing agent is one of the important factors influencing the electron density of the product.