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Bright red-emitting highly reliable styryl probe with large stokes shift for visualizing mitochondria in live cells under wash-free conditions
摘要： Bright red-emitting pyridinium cyanine based styryl probe 2 is synthesized in good yields. Probe 2 demonstrated a large Stokes’ shift (Δλ ≈ 128 nm, 4227 cm-1 in DCM) and excellent fluorescent quantum yield (?fl ≈ 0.2 - 0.7) due to strong Intra-molecular charge transfer (ICT). Probe 2 found to exhibit exceptional selectivity for cellular mitochondria in both normal (COS-7) and cancer (A549) cell lines. Probe 2 is readily applicable as a “wash-free” dye to visualize mitochondria as it does not require post-staining washing prior to imaging. Styryl probe 2 also showed an excellent biocompatibility as the calculated LC50 (lethal concentration, 50%) value was > 20 μM. Probe 2 emission did not show any interferences from anionic species or other biological molecules. Probe 2 is readily excitable (λex ~460 and λem ~618 nm) with the available laser (454 nm) in commercial microscopes and thus it can be a useful probe for mitochondrial tracking in live cells.
关键词： Wash-free application,Biocompatibility,Intra-molecular charge transfer (ICT),Styryl dye,Large Stokes shift,Mitochondria selectivity
更新于2025-09-23 15:23:52
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A pH-insensitive near-infrared fluorescent probe for wash-free lysosome-specific tracking with long time during physiological and pathological processes
摘要： A novel dicyanomethylene-4H-pyran based lysosome-specific near-infrared (NIR) probe, DCM-ML, is reported. It exhibits very weak fluorescence (Ф=0.007) in aqueous solution due to the energy relaxation from intramolecular rotation; but a large "off-on" NIR emission (20-folds) after specifically labeling lysosomes due to the local high viscosity instead of the local pH by both one- and two-photon excitation microscopy. Therefore, the imaging ability of DCM-ML will not confront the problems suffered by the currently used and reported pH-dependent lysosome probes (e.g., once lysosomal pH increases, their fluorescence gets quenched); The pH-independent feature of DCM-ML is further demonstrated to be a great advantage for tracing lysosomes’ movements in a relatively long time, even during cellular processes when lysosomes are suffered from external stimulations that induce pH increase and apoptosis. In addition to the characteristics of pH-insensitivity and "off-on" NIR emission, DCM-ML also possesses characteristics of excellent membrane permeability, high selectivity, low cytotoxicity, good photostability and large Stokes shift (> 200 nm), which qualify it as a superior wash-free NIR probe for tracking dynamic changes of lysosomes under both physiological and toxicological conditions. Hence, we believe DCM-ML may find a great potential of application in the future.
关键词： pH-insensitivity,Long-term tracking,Wash-free,"Off-on",Lysosome-specific imaging,Near-infrared
更新于2025-09-23 15:22:29