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应用DAB法对细胞色素氧化酶(CCO)和应用铁氰化物技术对琥珀酸脱氢酶(SDH)作超微结构定位,已被广泛接受。从理论上讲,SDH是定位于线粒体嵴膜的内侧面,CCO是定位于线粒体嵴膜的外侧面,但是从众多文献看,由于反应产物过溢及移位扩散,实际上均定位于线粒体嵴腔及线粒体外窒。 1.对SDE超微结构技术的改进:(1)对固定剂浓度及固定时间作合理筛选,以最大程度保存SDH的活性;(2)组织在反应基质中孵育后,再用DAB处理以增强反差;(3)对不同时间孵育的组织作动态观察,以选择最佳孵育时间,发现短期孵育后,SDH主要定位于线粒体嵴膜及内膜,当延长孵育时
It has been widely accepted that the DAB method for the cytosolic oxidase (CCO) and the application of the ferricyanide technology to succinate dehydrogenase (SDH) for ultra-structural localization. Theoretically, SDH is located on the inner side of the mitochondrial crest membrane, CCO is located on the outer side of the mitochondrial crest membrane, but from many literatures, due to the overflow of the reaction products and shift diffusion, are actually located in the mitochondrial crest Cavity and mitochondria outside the stifled. 1. Improve the ultrastructure of SDE technology: (1) Make reasonable screening of fixative concentration and fixed time to maximize the activity of SDH; (2) After the tissue is incubated in the reaction matrix and then treated with DAB to enhance Contrast; (3) Dynamic observation of tissue incubated at different times to select the best incubation time and found that after short-term incubation, SDH mainly located in mitochondrial ridge and endometrium, when extended incubation