The scientific literature on mitochondria has increased significantly over the years due to findings that these organelles have widespread roles in the onset and progression of pathological conditions such as metabolic disorders, neurodegenerative and cardiovascular diseases, inflammation, and cancer. Researchers have extensively explored how mitochondrial properties and functions are modified in different models, often using fluorescent inner mitochondrial membrane potential (ΔΨm) probes to assess functional mitochondrial aspects such as protonmotive force and oxidative phosphorylation. This review provides an overview of existing techniques to measure ΔpH and ΔΨm, highlighting their advantages, limitations, and applications. It discusses drawbacks of ΔΨm probes, especially when used without calibration, and conditions where alternative methods should replace ΔΨm measurements for the benefit of the specific scientific objectives entailed. Studies investigating mitochondria and their vast biological roles would be significantly advanced by the understanding of the correct applications as well as limitations of protonmotive force measurements and use of fluorescent ΔΨm probes, adopting more precise, artifact-free, sensitive, and quantitative measurements of mitochondrial functionality.

中文翻译：

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如何以及何时测量线粒体内膜电位

多年来，有关线粒体的科学文献显着增加，因为发现这些细胞器在代谢紊乱、神经退行性和心血管疾病、炎症和癌症等病理状况的发生和进展中发挥着广泛的作用。研究人员广泛探索了如何在不同模型中修改线粒体特性和功能，通常使用荧光线粒体内膜电位 (ΔΨm) 探针来评估线粒体功能，例如质子动力和氧化磷酸化。本综述概述了测量 ΔpH 和 ΔΨm 的现有技术，强调了它们的优点、局限性和应用。它讨论了 ΔΨm 探头的缺点，特别是在没有校准的情况下使用时，以及为了实现特定科学目标而应采用替代方法取代 ΔΨm 测量的条件。通过了解质子动力测量的正确应用和局限性以及荧光 ΔΨm 探针的使用，采用更精确、无伪影、灵敏和定量的线粒体功能测量，调查线粒体及其广泛生物学作用的研究将得到显着推进。