Buravkov S V, Pogodina M V, Buravkova L B
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University, Moscow, Russia,
Bull Exp Biol Med. 2014 Sep;157(5):654-8. doi: 10.1007/s10517-014-2637-3. Epub 2014 Sep 26.
Specific fluorescent mitochondrial dyes JC-1, MitoTracker Red FM, and MitoTracker Red CMXRos were studied on cultures of mesenchymal stromal stem cells from human adipose tissue. Static and dynamic experiments demonstrated the dependence of dye accumulation in the mitochondria and cytoplasm on mitochondrial potential in the presence of oxidative phosphorylation uncoupler CCCP. JC-1 dye demonstrated most pronounced bleaching upon repeated laser exposure even in very low doses, which should be taken into account when planning dynamic experiments on live cells. Moreover, this dye requires two-wave recording in red and green spectral bands. MitoTracker Red FM and MitoTracker Red CMXRos are resistant to bleaching, but for the latter, the intensity of fluorescence reflects not only the mitochondrial potential. MitoTracker Red CMXRos covalently binds to sulfhydryl groups in mitochondrial lipids and proteins, which can lead to misinterpretation of the fluorescence intensity-mitochondrial potential dependence.
对人脂肪组织间充质基质干细胞培养物研究了特异性荧光线粒体染料JC-1、MitoTracker Red FM和MitoTracker Red CMXRos。静态和动态实验表明，在存在氧化磷酸化解偶联剂CCCP的情况下，染料在线粒体和细胞质中的积累取决于线粒体电位。即使在非常低的剂量下，JC-1染料在重复激光照射时也表现出最明显的漂白现象，在规划活细胞动态实验时应考虑到这一点。此外，这种染料需要在红色和绿色光谱带进行双波记录。MitoTracker Red FM和MitoTracker Red CMXRos抗漂白，但对于后者，荧光强度不仅反映线粒体电位。MitoTracker Red CMXRos与线粒体脂质和蛋白质中的巯基共价结合，这可能导致对荧光强度-线粒体电位依赖性的错误解读。
