An ability to visualize HCN in mitochondria in real time may permit additional insights into the critical toxicological and physiological roles this classic toxin plays in living organisms. Herein, we report a mitochondria-specific coumarin pyrrolidinium-derived fluorescence probe (MRP1) that permits the real-time ratiometric imaging of HCN in living cells. The response is specific, sensitive (detection limit is ca. 65.6 nM), rapid (within 1 s), and reversible. Probe MRP1 contains a benzyl chloride subunit designed to enhance retention within the mitochondria under conditions where the mitochondria membrane potential is eliminated. It has proved effective in visualizing different concentrations of exogenous HCN in the mitochondria of HepG2 cells, as well as the imaging of endogenous HCN in the mitochondria of PC12 cells and within neurons. Fluctuations in HCN levels arising from the intracellular generation of HCN could be readily detected.

Labile Fe2+ has been considered to be a metabolically active and regulatory form of cellular iron. Monitoring the dynamic level of labile Fe2+ in biological systems is vital for evaluating the iron related biological processes and diseases as well as dissecting the exact physiological and pathophysiological functions of the labile Fe2+. Herein, we rationally constructed a coumarin-based fluorescent probe for sensing labile Fe2+ in living systems based on a novel Fe2+ meditated cyclization reaction strategy. The probe showed a highly selective and sensitive response to Fe2+, and the detection limit was determined to be 45 nM. Significantly, the probe displayed fast response to Fe2+, with the sensing reaction completed in 2 min, which is beneficial for real time sensing. The application of the probe for sensing different concentrations of labile Fe2+ in living cells has been conducted. In addition, the basal and endogenous levels of labile Fe2+ in living systems were also successfully monitored.

Thiols and primary aliphatic amines (PAA) are ubiquitous and extremely important species in biological systems. They perform significant interplaying roles in complex biological events. A single fluorescent probe differentiating both thiols and PAA can contribute to understanding the intrinsic inter-relationship of thiols and PAA in biological processes. Herein, we rationally constructed the first fluorescent probe that can respond to thiols and PAA in different fluorescence channels. The probe exhibited a high selectivity and sensitivity to thiols and PAA. In addition, it displayed sequential sensing ability when the thiols and PAA coexisted. The application experiments indicated that the probe can be used for sensing thiols and PAA in human blood serum. Moreover, the fluorescence imaging of endogenous thiols and PAA as well as antihypertensive drugs captopril and amlodipine in living cells were successfully conducted.