A facile click reaction to fabricate a FRET-based ratiometric fluorescent Cu2+ probe

A facile one-step Cu(I)-catalyzed click reaction, between a dansyl-azide and a propargyl-substituted rhodamine B hydrazide, is employed to fabricate a novel FRET ratiometric off-on fluorescent probe. The sensitive emission of the donor, a dansyl group, overlaps perfectly with the absorption of the acceptor, xanthene in the open-ring rhodamine. The proposed probe shows high selectivity towards Cu2+. The ratio of emission intensities at 568 and 540 nm (I-568/I-540) exhibits a drastic 28-fold enhancement upon addition of Cu2+. The probe shows an excellent linear relationship between emission ratios and the concentrations of Cu2+ from 10 to 50 mu M, with a detection limit (S/N = 3) of 0.12 mu M. The preliminary cellular studies demonstrated that the probe is cell membrane permeable and could be applied for ratiometric fluorescence imaging of intracellular Cu2+ with almost no cytotoxicity. The ingenuity of the probe design is to construct a FRET donor-acceptor interconnector and a selective receptor simultaneously by click reaction. The strategy was verified to have great potential for developing novel FRET probes for Cu2+.