Hynes James, Swiss Rachel L, Will Yvonne
Luxcel Biosciences, BioInnovation Centre, UCC, Cork, Ireland.
Methods Mol Biol. 2012;810:59-72. doi: 10.1007/978-1-61779-382-0_4.
Interest in the investigation of mitochondrial dysfunction has seen a resurgence over recent years due to the implication of such dysfunction in both drug-induced toxicity and a variety of disease states. Here, we describe a methodology to assist in such investigations whereby the oxygen consumption of isolated mitochondria is assessed in a high-throughput fashion using a phosphorescent oxygen-sensitive probe, standard microtitre plates, and plate reader detection. The protocols provided describe the required isolation procedures, initial assay optimization, and subsequent compound screening. Typical data is also provided illustrating the expected activity levels as well as recommended plate maps and data analysis approaches.
近年来，由于线粒体功能障碍与药物诱导的毒性以及多种疾病状态有关，对其研究的兴趣再度兴起。在此，我们描述一种有助于此类研究的方法，即使用磷光氧敏探针、标准微孔板和平板读数器检测，以高通量方式评估分离线粒体的氧消耗。所提供的方案描述了所需的分离程序、初始测定优化以及后续的化合物筛选。还提供了典型数据，说明了预期的活性水平以及推荐的板图和数据分析方法。
