Tetramethyl rhodamine methyl ester (TMRM) is suitable for cytofluorometric measurements of mitochondrial membrane potential in cells treated with digitonin
Tetramethyl rhodamine methyl ester (TMRM) is suitable for cytofluorometric measurements of mitochondrial membrane potential in cells treated with digitonin
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DOI：
10.1023/a:1020193906974
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发表时间：
1999-02-01
影响因子：
4
通讯作者：
Houstek, J
中科院分区：
文献类型：
--
作者：
Floryk, D;Houstek, J
以TMRM为阳离子线粒体选择性探针，建立了一种细胞荧光分析线粒体膜电位的新方法。该方法基于用毛地黄皂苷有限处理培养的细胞，毛地黄皂苷使质膜透化并使线粒体完整。因此，TMRM染色细胞的所得信号仅代表线粒体中积累的探针。以成纤维细胞和胞质杂交细胞为模型细胞系统，描述了毛地黄皂苷处理和TMRM染色的最佳条件：用缬氨霉素、KCN和抗霉素A破坏TMRM信号，用FCCP滴定逐渐降低Δ Psi并破坏Δ Psi的稳定性。该方法适用于不同类型培养细胞中Δ Psi的灵敏测量。
A new method for cytofluorometric analysis of mitochondrial membrane potential Delta Psi 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 Delta Psi and characterise the stability of Delta Psi. The method is suitable for sensitive measurement of Delta Psi in different types of cultured cells.