线粒体是一种具有双层膜结构的细胞器,参与细胞新陈代谢过程的能量循环以及离子平衡过程,在细胞生理过程中具有极其重要的意义.一些小分子荧光染料/探针结构中带有正电荷,因受到线粒体内膜负电势的牵引而标记在线粒体上,为研究线粒体的形态或功能提供了重要的可视化成像工具.然而,大多数线粒体染料/探针对线粒体的靶向标记稳定性仍不够理想,因为线粒体电势处于不断的动态变化中,当电势降低时,对染料的亲和力相应降低.尤其在病理条件下(比如细胞凋亡)细胞代谢受到阻滞时,线粒体膜电势显著降低,阳离子染料将扩散离开线粒体,造成非特异性荧光.最近,Kim团队和本人课题组提出可固定线粒体探针的新概念,用活性基团将荧光分子探针通过共价键固定在线粒体中,开发了稳定靶向线粒体中的定量探测微环境pH值、粘度、膜电势荧光探针.我们认为,对于追踪和探测具有高度动态变化特性的线粒体而言,开发可固定的线粒体荧光分子探针是必然趋势,因此本文进行评述和展望.%Mitochondria, a cell organelle with a double membrane structure, plays significant roles in cell metabolism, including energy circulation and ions balance.Some small positively charged molecule fluorescent dyes/probes can label mitochondria due to the traction of the mitochondrial inner membrane negative potential, providing an important visual imaging tool for investigation of mitochondrial morphology and function.However, most mitochondrial dyes/probes are unsatisfactory as targeting marker of mitochondria , because the mitochondrial potential is constantly changing, when the potential reduced, the affinity of the dye accordingly decreases.Especially, the mitochondrial membrane potential will be significantly reduced when cell metabolism is blocked in pathological conditions( such as apoptosis) , thus cationic dyes will spread away from mitochondria and result in non-specific fluorescence.Recently, the Kim team and our team proposed a new concept of fixed mitochondrial probe, with the active group of fluorescent molecular probe can be fixed in the mitochondria through the covalent bond, and developed a series of fluorescent probes that stably targeted mitochondria for the quantitative detection of microenvironment pH, viscosity, and membrane potential.We believe that the development of immobilized mitochondrial fluorescent molecular probes is an inevitable trend in tracking and detecting mitochondria with highly dynamic characteristics.Therefore, this paper reviews and prospects the fixable fluorescent dyes of mitochondria.
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