We evaluated the ability of different fluorescent indicators by various analytical instruments, including a laser scanning confocal microscope (LSCM), fluorescence plate reader, and flow cytometer (FCM), to measure the mitochondrial membrane potential (ΔΨm) of cardiac H9c2 cells during oxidative stress-induced mitochondrial injury. The mitochondrial oxygen consumption rate and a transmission electron microscope were used to detect changes in mitochondrial functions and morphology, respectively. Cardiac H9c2 cells were exposed to H₂O₂ (500, 750, 1000, and 1250 μM) to induce mitochondrial oxidative stress injury, and fluorescent indicators including tetramethyl rhodamine ethyl ester (TMRE), 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolocarbocyanine iodide (JC-1), and rhodamine 123 (R123) were used to detect changes in ΔΨm using an LSCM, fluorescence plate reader, and FCM. The decrease in ΔΨm caused by H₂O₂ was determined by endpoint and dynamic analyses after staining with JC-1 or TMRE. With the R123 probe, the LSCM could only detect the change in ΔΨm caused by 1000 μM H₂O₂. Moreover, R123 was less effective than JC-1 and TMRE for measurement of ΔΨm by the LSCM. Our data indicated that an LSCM is the most suitable instrument to detect dynamic changes in ΔΨm, whereas all three instruments can detect ΔΨm at the endpoint.

我们通过各种分析仪器（包括激光扫描共聚焦显微镜（LSCM），荧光板读取器和流式细胞仪（FCM））评估了不同荧光指示剂在氧化应激期间测量心脏H9c2细胞的线粒体膜电位（ΔΨm）的能力。引起的线粒体损伤。线粒体耗氧率和透射电镜分别用于检测线粒体功能和形态的变化。心脏H9c2细胞暴露于H ₂ O ₂（500、750、1000和1250μM）诱导线粒体氧化应激损伤，荧光指示剂包括四甲基若丹明乙酯（TMRE），5,5'，6,6'-tetrachloro-1,1'，3,3使用LSCM，荧光板读数器和FCM，使用'-四乙基苯并咪唑并羰基花青碘化物（JC-1）和若丹明123（R123）来检测ΔΨm的变化。由H ₂ O ₂引起的ΔΨm降低是通过用JC-1或TMRE染色后的终点分析和动态分析确定的。使用R123探针，LSCM只能检测到1000μMH ₂ O ₂引起的ΔΨm变化。而且，R123在通过LSCM测量ΔΨm方面不如JC-1和TMRE有效。我们的数据表明，LSCM是最适合检测Δdynamicm动态变化的仪器，而所有三种仪器都可以在终点检测ΔΨm。