Floryk D, Houstĕk J
Institute of Physiology, Academy of Sciences of the Czech Republic, Prague.
Biosci Rep. 1999 Feb;19(1):27-34. doi: 10.1023/a:1020193906974.
A new method for cytofluorometric analysis of mitochondrial membrane potential deltapsi has been developed by using TMRM as a cationic, mitochondrial selective probe. The method is based on limited treatment of cultured cells with digitonin which permeabilises the plasma membrane and leaves mitochondria intact. The resulting signal of TMRM-stained cells thus represents only the probe accumulated in mitochondria. Fibroblasts and cybrids were used as a model cell systems and optimal conditions for digitonin treatment and staining by TMRM were described. The TMRM signal collapsed by valinomycin, KCN and antimycin A and FCCP titration was used to gradually lower deltapsi and characterise the stability of deltapsi. The method is suitable for sensitive measurement of deltapsi in different types of cultured cells.
通过使用四甲基罗丹明甲酯（TMRM）作为阳离子线粒体选择性探针，已开发出一种用于细胞荧光分析线粒体膜电位（Δψ）的新方法。该方法基于用洋地黄皂苷对培养细胞进行有限处理，洋地黄皂苷可使质膜通透，而线粒体保持完整。因此，TMRM染色细胞产生的信号仅代表积累在线粒体中的探针。成纤维细胞和胞质杂种用作模型细胞系统，并描述了洋地黄皂苷处理和TMRM染色的最佳条件。缬氨霉素、氰化钾、抗霉素A使TMRM信号消失，FCCP滴定用于逐渐降低Δψ并表征Δψ的稳定性。该方法适用于灵敏测量不同类型培养细胞中的Δψ。
