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Updated: May 20, 2026
Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
Published on: January 24, 2017
A near-infrared multifunctional fluorescent probe for simultaneous tracking of mitochondrial viscosity/polarity and
Can Liu1, Tong Han1, Xiao-Ran Yang1
1College of Laboratory Medicine, Institute of Pathogen Biology and Immunology, Hebei Key Laboratory of Neuropharmacology, Hebei Key Laboratory of Quality & Safety Analysis-Testing for Agro-Products and Food and Zhang Jiakou Key Laboratory of Organic Light Functional Materials, Hebei North University, Zhangjiakou, Hebei Province, 075000, China.
Abstract:
Mitochondrial microenvironment abnormalities (viscosity, polarity, and ONOO-) are closely linked to diseases, but near-infrared (NIR) probes for simultaneous detection of three parameters remain scarce. Here, we rationally designed a NIR multifunctional probe (PEQ-Bf) featuring a double bond linked N,N-dimethylamino and pyridinium quaternary salt. PEQ-Bf enabled visualization viscosity/polarity and ONOO- at 650 nm and 500 nm, respectively, thereby avoiding signal crosstalk. And PEQ-Bf exhibited high sensitivity (LOD = 80.2 nM), a wide linear range (0-14 μM), excellent selectivity, and strong anti-interference capability toward ONOO-. Meanwhile PEQ-Bf successfully visualized viscosity increase, polarity decrease, and ONOO- elevation in mitochondria under high-glucose conditions. Using this probe, we revealed for the first time that gliclazide and resveratrol alleviated diabetes by inhibiting oxidative stress related mitochondrial microenvironment deterioration. Furthermore, PEQ-Bf enabled in situ imaging of viscosity and polarity changes in diabetic and chronic wound-healing mouse models. Thus, PEQ-Bf offered a promising chemical tool for elucidating the role of mitochondrial microenvironment in disease progression.
