Division of Molecular Medicine, Ruđer Bošković Institute, Zagreb, Croatia. INSERM U1124 University Paris Descartes, Paris, France.
Methods Appl Fluoresc. 2017 Mar 22;5(1):015007. doi: 10.1088/2050-6120/aa5e64.
We evaluated our phosphonium-based fluorescent probes for selective staining of mitochondria. Currently used probes for monitoring mitochondrial membrane potential show varying degrees of interference with cell metabolism, photo-induced damage and probe binding. Here presented probes are characterised by highly efficient cellular uptake and specific accumulation in mitochondria. Fluorescent detection of the probes was accomplished using flow cytometry and confocal microscopy imaging of yeast and mammalian cells. Toxicity analysis (impedimetry-xCELLigence for the cellular proliferation and Seahorse technology for respiratory properties) confirms that these dyes exhibit no-toxicity on mitochondrial or cellular functioning even for long time incubation. The excellent chemical and photophysical stability of the dyes makes them promising leads toward improved fluorescent probes. Therefore, the probes described here offer to circumvent the problems associated with existing-probe's limitations.
我们评估了基于鏻的荧光探针在选择性染色线粒体中的应用。目前用于监测线粒体膜电位的探针在不同程度上存在与细胞代谢、光诱导损伤和探针结合的干扰。本研究中提出的探针具有高效的细胞摄取能力和特异性在线粒体中的积累。通过流式细胞术和酵母及哺乳动物细胞的共聚焦显微镜成像来实现探针的荧光检测。毒性分析（细胞增殖的阻抗检测-xCELLigence 和呼吸特性的 Seahorse 技术）证实，即使长时间孵育，这些染料对线粒体或细胞功能也没有毒性。染料具有优异的化学和光物理稳定性，这使它们成为改进荧光探针的有前途的先导化合物。因此，本文描述的探针为克服现有探针的局限性提供了一种方法。
