New method to assess mitophagy flux by flow cytometry
New method to assess mitophagy flux by flow cytometry
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DOI：
10.1080/15548627.2015.1034403
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发表时间：
2015-05-01
期刊：
影响因子：
13.3
通讯作者：
Boya, Patricia
中科院分区：
文献类型：
--
作者：
Mauro-Lizcano, Marta;Esteban-Martinez, Lorena;Boya, Patricia
线粒体自噬，也称为线粒体自噬，是一种基于自噬体的线粒体降解过程，可在细胞应激后消除不需要或受损的线粒体。大多数涉及线粒体自噬的研究依赖于荧光显微镜对线粒体-自噬体共定位的分析。然而，鉴于线粒体自噬在生物体生理学和病理学中的基本作用，迫切需要新的定量方法来研究这一过程。在这里，我们描述了一种基于流式细胞术的方法，通过使用 MitoTracker Deep Red（一种广泛使用的线粒体选择性探针）来确定线粒体自噬。与选择性抑制剂结合使用，可以测定线粒体自噬通量。在这里，我们测试了在细胞系以及原代细胞和组织培养物中使用该方法的有效性。
Mitochondrial autophagy, also known as mitophagy, is an autophagosome-based mitochondrial degradation process that eliminates unwanted or damaged mitochondria after cell stress. Most studies dealing with mitophagy rely on the analysis by fluorescence microscopy of mitochondrial-autophagosome colocalization. However, given the fundamental role of mitophagy in the physiology and pathology of organisms, there is an urgent need for novel quantitative methods with which to study this process. Here, we describe a flow cytometry-based approach to determine mitophagy by using MitoTracker Deep Red, a widely used mitochondria-selective probe. Used in combination with selective inhibitors it may allow for the determination of mitophagy flux. Here, we test the validity of the use of this method in cell lines and in primary cell and tissue cultures.