Live cells imaging using a turn-on FRET-based BODIPY probe for biothiols

We designed a red-emitting turn-on FRET-based molecular probe 1 for selective detection of cysteine and homocysteine. Probe 1 shows significant fluorescence enhancement after cleavage of the 2, 4-dinitrobenzensulfonyl (DNBS) unit from the fluorophore upon thiols treatment. The precursor of probe 1, BNM153, is a moderate quantum yield FRET dye which contributes a minimum emission leakage from its donor part. We synthesized this assembly by connecting a low quantum yield (less than 1%) BODIPY donor to a high quantum yield BODIPY acceptor via a 1, 3-triazine bridge system. It is noteworthy that the majority of the non-radiative energy loss of donor (BDN) was converted to the acceptor (BDM)'s fluorescence output with minimum leaks of donor emission. The fluorescence sensing mechanism of probe 1 was illustrated by fluorescence spectroscopy, kinetic measurements, HPLC-MS analysis and DFT calculations. Probe 1 is pH-independent at the physiological pH range. Finally, live cells imaging demonstrated the utility of probe 1 as a biosensor for thiols. (C) 2014 Elsevier Ltd. All rights reserved.