Abstract

Real-time imaging and monitoring of biothiols in living cells are essential for understanding pathophysiological processes. However, the design of the fluorescent probe that has accurate and repeatable real-time monitoring capabilities for these targets is highly challenging. In this study, we prepared a fluorescent sensor, Lc-NBD-Cu(II), which contains a N1, N1, N2-tris-(pyridin-2-ylmethyl) ethane-1,2-diamine as a Cu(II) chelating unit and a 7-nitrobenz-2-oxa-1,3-diazole fluorophore to detect Cysteine (Cys). Emission changes promoted by addition of Cys to this probe are distinctive and correspond to a range of processes including Cys induced loss of Cu(II) from Lc-NBD-Cu(II) to form Lc-NBD, Cu(I) oxidation to reform Cu(II), Cys oxidation to form Cys-Cys, Cu(II) binding to Lc-NBD to reform Lc-NBD-Cu(II), and competitive binding of Cu(II) to Cys-Cys. The study also shows that Lc-NBD-Cu(II) maintains high stability during the sensing process and that it can be utilized over a number of detection cycles. Finally, the findings show that Lc-NBD-Cu(II) can be utilized to repetitively sense Cys in living HeLa cells.

Full Text
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