Department of Biochemistry, Faculty of Medicine, University of Toronto, Toronto, Ontario, M5S 1A8, Canada.
Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, Institute for Biomedical and Biomaterials Engineering, Departments of Biochemistry and Chemistry, University of Toronto, Toronto, Ontario, M5S 1A8, Canada.
Angew Chem Int Ed Engl. 2018 Jul 16;57(29):8891-8895. doi: 10.1002/anie.201802796. Epub 2018 Jun 19.
The measurement of physicochemical parameters in living cells can provide information on individual cellular organelles, helping us to understand subcellular function in health and disease. While organelle-specific chemical probes have allowed qualitative evaluation of microenvironmental variations, the simultaneous quantification of mitochondrial local microviscosity (η ) and micropolarity (ϵ ), along with concurrent structural variations, has remained an unmet need. Herein, we describe a new multifunctional mitochondrial probe (MMP) for simultaneous monitoring of η and ϵ by fluorescence lifetime and emission intensity recordings, respectively. The MMP enables highly precise measurements of η and ϵ in the presence of a variety of agents perturbing cellular function, and the observed changes can also be correlated with alterations in mitochondrial network morphology and motility. This strategy represents a promising tool for the analysis of subtle changes in organellar structure.
活细胞中理化参数的测量可以提供有关单个细胞细胞器的信息,有助于我们了解健康和疾病状态下的亚细胞功能。虽然细胞器特异性化学探针已允许对微环境变化进行定性评估,但同时定量线粒体局部微粘度(η)和微极性(ϵ),以及同时进行结构变化,仍然是一个未满足的需求。在这里,我们描述了一种新的多功能线粒体探针(MMP),可分别通过荧光寿命和发射强度记录来同时监测η和ϵ。MMP 能够在各种干扰细胞功能的试剂存在下进行高度精确的 η 和 ϵ 测量,并且观察到的变化也可以与线粒体网络形态和运动的改变相关联。该策略代表了分析细胞器结构细微变化的有前途的工具。