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  • Title: A Copper Nanocluster-Based Fluorescent Probe for Real-Time Imaging and Ratiometric Biosensing of Calcium Ions in Neurons.
    Author: Liu Z, Jing X, Zhang S, Tian Y.
    Journal: Anal Chem; 2019 Feb 05; 91(3):2488-2497. PubMed ID: 30618245.
    Abstract:
    Fluorescent calcium ion (Ca2+) sensing and imaging have become an essential technique for investigation of signaling pathways of Ca2+ and understanding the role of Ca2+ in neurodegenerative disease. Herein a copper nanocluster (CuNC)-based ratiometric fluorescent probe was developed for real-time sensing and imaging of Ca2+ in neurons, in which a specific Ca2+ ligand with two formaldehyde groups was synthesized and further conjugated with polyethylenimine (PEI) to form a new ligand molecule for the synthesis of CuNCs. Meanwhile, water-soluble Alex Fluor 660 NHS ester was immobilized onto CuNCs as a reference element. The developed ratiometric fluorescence nanoprobe demonstrated a good linearity with Ca2+ concentration in the range of 2-350 μM, and a detection limit down to 220 ± 11 nM was achieved. In addition, the response time of the present probe for Ca2+ was found to be less than 2 s with good stability and high selectivity. Taking advantage of the low cytotoxicity and good biocompatibility of the developed nanoprobe, it was discovered that the histamine-induced cytoplasmic Ca2+ increase in various parts of neurons was different. Moreover, it was found O2•--induced cytoplasmic Ca2+ burst and O2•--induced neuronal death possibly resulted from Ca2+ overload in the neurons.
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