Two new sulfur-containing ferrocenylimidazo [4,5-b]pyridines, 2-ferrocenyl-5-(thiophen-2-yl)-1H-imidazo [4,5-b]pyridine (L1) and 2-ferrocenyl-5-(thiophen-3-yl)-1H-imidazo[4,5-b]pyridine (L2), were synthesized and characterized. L1 and L2 behaved as highly selective fluorescent "turn-off", redox (ΔE 1/2 = 0.25 V for L1 and ΔE 1/2 = 0.22 V for L2) and UV active receptors for Hg 2 + ions through the formation of the corresponding 2 : 1 and 1 : 1 (receptor:metal ion) complexes, with detection limits 4.47 × 10 À 8 M and 2.21 × 10 À 7 M, respectively. Furthermore, the quenched fluorescence of L1 was recovered on iodide ion addition. The sensing mechanism study suggested that although the imidazo[4,5-b]pyridine is the main coordination unit providing N, N binding sites, the added thienyls have improved the fluorescence emission and Hg 2 + sensing selectivity of L1 and L2, and obviously influenced the coordination mode. Finally, living cell imaging and test strip experiments demonstrated the potential application of L1 in fluorescent visualization of Hg 2 + ions in biological and environmental systems.