Sivandzade Farzane, Bhalerao Aditya, Cucullo Luca
Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, Amarillo, TX 79106, USA.
Center for Blood-Brain Barrier Research, Texas Tech University Health Sciences Center, Amarillo, TX 79106, USA.
Bio Protoc. 2019 Jan 5;9(1). doi: 10.21769/BioProtoc.3128.
In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the results across laboratories strongly depends upon following well validated and reliable protocols along with the appropriate controls. Herein, we provide a practical user guide for monitoring mitochondrial membrane potential in whole cells using a fluorescent cationic probe. The data analysis of mitochondrial membrane potential we provide is one associated with the impact of xenobiotics such as tobacco smoking on blood-brain barrier endothelial cells including both mouse primary (mBMEC) and a mouse-based endothelial cell line (bEnd3) in a side by side comparison.
近年来,荧光染料经常被用于监测线粒体膜电位,以评估线粒体的活力和功能。然而,不同实验室结果的可重复性在很大程度上取决于遵循经过充分验证且可靠的方案以及适当的对照。在此,我们提供一份实用的用户指南,介绍如何使用荧光阳离子探针监测全细胞中的线粒体膜电位。我们提供的线粒体膜电位数据分析是一项关于诸如吸烟等外源性物质对血脑屏障内皮细胞影响的分析,其中包括小鼠原代脑微血管内皮细胞(mBMEC)和一种基于小鼠的内皮细胞系(bEnd3),并进行了并排比较。