Acosta D, Wenzel D G
Histochem J. 1975 Jan;7(1):45-56. doi: 10.1007/BF01004831.
A cytochemical permeability test for the detection of injury to in situ mitochondria of cultured heart cells is presented. The test is based on the increased rate at which injured mitochondria stain for succinate dehydrogenase activity. Whereas an intact inner mitochondrial membrane limits the rate at which Nitro Blue tetrazolium and phenazine methosulphate reach succinate dehydrogenase, injured mitochondria allow these reactants to reach the enzyme more rapidly to form microscopically-observable formazan granules. The extent of staining at fixed durations of incubation with the reactants was assessed on a blind basis with pseudo dark-field microscopy, using a standardized rating scale. Differences in the staining of control and treated cells were analysed statistically by a semi-quantitative method. Treatment of the cultures with either vitamin A or chlorpromazine, resulted in more rapid mitochondrial staining. Brief pre-fixation of the cells with cold acetone also labilized the mitochondria as did a delay in the change of culture medium.
本文介绍了一种用于检测培养的心脏细胞原位线粒体损伤的细胞化学通透性试验。该试验基于受损线粒体琥珀酸脱氢酶活性染色速率的增加。完整的线粒体内膜会限制硝基蓝四唑和吩嗪硫酸甲酯与琥珀酸脱氢酶接触的速率，而受损的线粒体则允许这些反应物更快地接触到该酶，从而形成显微镜下可观察到的甲臜颗粒。使用标准化评分量表，通过伪暗视野显微镜在盲法基础上评估与反应物孵育固定时间后的染色程度。采用半定量方法对对照细胞和处理细胞的染色差异进行统计学分析。用维生素A或氯丙嗪处理培养物，会导致线粒体染色更快。用冷丙酮对细胞进行短暂预固定，以及延迟更换培养基，也会使线粒体变得不稳定。
