Detection and imaging of Carbon monoxide releasing Molecule-2 in HeLa cells and zebrafish using a Metal-Free...

By incorporating 4-nitrobenzene as the recognition unit into a stable hemicyanine skeleton based on IR 780, a designed near-infrared fluorescent off-on probe DNXI shows excellent analytical performance in real-time monitoring of carbon monoxide releasing molecule-2 (CORM-2) in living HeLa cells and zebrafish without transition metals. The response mechanism of CORM-2 with DNXI may reduce the nitro group to an amino group, followed by 1,6-rearrangement elimination reaction, resulting in the recovery of both color and fluorescence signal. As a result, DNXI shows very low background signal, which is rather desired for achieving sensitive detection of CORM-2. Compared with the existing CORM-2 probes, DNXI shows excellent optical performance in vitro and in vivo, high selectivity and sensitivity to CORM-2, as well as near-infrared fluorescence emission 712 nm, with a low detection limit of 103 nM. More importantly, DNXI is low cytotoxic, cell membrane permeable, and its applicability has been demonstrated for monitoring CORM-2 in living HeLa cells and zebrafish. These superior properties of the probe enable it have great potential to be used in biological systems or in vivo related studies.