摘要
一、琥珀酸脱氢酶(SDH)超微结构技术的改进:合理筛选固定时间及固定剂浓度;组织在基质中孵育后再经DAB处理,对不同时间孵育后的组织作动态性观察。二、细胞色素氧化酶(CCO):选择合适的固定剂种类,固定时间及固定剂浓度;筛选最适孵育时间;低温脱水。
In order to prevent the excessive reaction product from entering the intracristate spaces and the outer compartment between the inner and outer limiting mitochondrial membrance we have established a reliable method for the localization of the cytochrome oxidase and succinic dehydrogenase. The results indicated that the better ultrastructural localization of cytochrome oxidase in the mitochondria of heart muscle from Wistar rat and excellent morphologic preservation were afforded by immersing the blocks in cold 2% formaldehyde for 10 min., incubating the blocks in the reaction medium added polyvinylpyrrolidone, carrying out dehydration process at the temperature of 4℃ Cytochrome oxidase was located on the cristal and inner membrances of the mitochondria after incu- bation for 45, 60, 90 min.. Fairly good results of succinic dehydrogenase have been obtained by fixing the blocks in cold 2% formaldehyde for 2 min., adding dimethyl sulfoxide (DMSO) to the incubation medium, and treating the blocks with 3,3'-diaminobenzidine (DAB) to an osmiophilic reaction product after the incubation. The results also showed that succinic dehydrogenase was located premialy on the cristal and inner mem-brances, then the reaction product filled intracristate spaces and outer compartment between inner and outer limiting mitochondrial membrances after incubation for a longer time.
出处
《解剖学杂志》
CAS
CSCD
北大核心
1989年第1期33-37,共5页
Chinese Journal of Anatomy
关键词
线粒体
结构定位
氧化酶
脱氢酶
Cytochrome oxidase, succinic dehydrogenase, mitochondria
