Creed Sarah, McKenzie Matthew
Monash Micro Imaging, Hudson Institute of Medical Research, Clayton, VIC, Australia.
Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia.
Methods Mol Biol. 2019;1928:69-76. doi: 10.1007/978-1-4939-9027-6_5.
The mitochondrial membrane potential (Δψ) drives the generation of ATP by mitochondria. Interestingly, Δψ is higher in many cancer cells comparted to healthy noncancerous cell types, providing a unique metabolic marker. This feature has also been exploited for therapeutic use by utilizing drugs that specifically accumulate in the mitochondria of cancer cells with high Δψ. As such, the assessment of Δψ can provide very useful information as to the metabolic state of a cancer cell, as well as its potential for malignancy. In addition, the measurement of Δψ can also be used to test the ability of novel anticancer therapies to disrupt mitochondrial metabolism and cause cell death.Here, we outline two methods for assessing Δψ in cancer cells using confocal microscopy and the potentiometric fluorescent dye tetramethylrhodamine methyl ester (TMRM). In the first protocol, we describe a technique to quantitatively measure Δψ, which can be used to compare Δψ between different cell types. In the second protocol, we describe a technique for assessing changes to Δψ over time, which can be used to determine the effectiveness of different therapeutic compounds or drugs in modulating mitochondrial function.
线粒体膜电位（Δψ）驱动线粒体产生ATP。有趣的是，与健康的非癌细胞类型相比，许多癌细胞中的Δψ更高，这提供了一种独特的代谢标志物。通过利用特异性积聚在具有高Δψ的癌细胞线粒体中的药物，这一特性也被用于治疗用途。因此，对Δψ的评估可以提供有关癌细胞代谢状态及其恶性潜能的非常有用的信息。此外，Δψ的测量还可用于测试新型抗癌疗法破坏线粒体代谢并导致细胞死亡的能力。在此，我们概述了两种使用共聚焦显微镜和电位荧光染料四甲基罗丹明甲酯（TMRM）评估癌细胞中Δψ的方法。在第一个方案中，我们描述了一种定量测量Δψ的技术，可用于比较不同细胞类型之间的Δψ。在第二个方案中，我们描述了一种评估Δψ随时间变化的技术，可用于确定不同治疗化合物或药物调节线粒体功能的有效性。
