2 research outputs found

    Chromis-1, a Ratiometric Fluorescent Probe Optimized for Two-Photon Microscopy Reveals Dynamic Changes in Labile Zn(II) in Differentiating Oligodendrocytes

    No full text
    Despite the significant advantages of two-photon excitation microscopy (TPEM) over traditional confocal fluorescence microscopy in live-cell imaging applications, including reduced phototoxicity and photobleaching, increased depth penetration, and minimized autofluorescence, only a few metal ion-selective fluorescent probes have been designed and optimized specifically for this technique. Building upon a donorā€“acceptor fluorophore architecture, we developed a membrane-permeant, ZnĀ­(II)-selective fluorescent probe, chromis-1, that exhibits a balanced two-photon cross section between its free and ZnĀ­(II)-bound form and responds with a large spectral shift suitable for emission-ratiometric imaging. With a <i>K</i><sub>d</sub> of 1.5 nM and wide dynamic range, the probe is well suited for visualizing temporal changes in buffered ZnĀ­(II) levels in live cells as demonstrated with mouse fibroblast cell cultures. Moreover, given the importance of zinc in the physiology and pathophysiology of the brain, we employed chromis-1 to monitor cytoplasmic concentrations of labile ZnĀ­(II) in oligodendrocytes, an important cellular constituent of the brain, at different stages of development in cell culture. These studies revealed a decrease in probe saturation upon differentiation to mature oligodendrocytes, implying significant changes to cellular zinc homeostasis during maturation with an overall reduction in cellular zinc availability. Optimized for TPEM, chromis-1 is especially well-suited for exploring the role of labile zinc pools in live cells under a broad range of physiological and pathological conditions

    Chromis-1, a Ratiometric Fluorescent Probe Optimized for Two-Photon Microscopy Reveals Dynamic Changes in Labile Zn(II) in Differentiating Oligodendrocytes

    No full text
    Despite the significant advantages of two-photon excitation microscopy (TPEM) over traditional confocal fluorescence microscopy in live-cell imaging applications, including reduced phototoxicity and photobleaching, increased depth penetration, and minimized autofluorescence, only a few metal ion-selective fluorescent probes have been designed and optimized specifically for this technique. Building upon a donorā€“acceptor fluorophore architecture, we developed a membrane-permeant, ZnĀ­(II)-selective fluorescent probe, chromis-1, that exhibits a balanced two-photon cross section between its free and ZnĀ­(II)-bound form and responds with a large spectral shift suitable for emission-ratiometric imaging. With a <i>K</i><sub>d</sub> of 1.5 nM and wide dynamic range, the probe is well suited for visualizing temporal changes in buffered ZnĀ­(II) levels in live cells as demonstrated with mouse fibroblast cell cultures. Moreover, given the importance of zinc in the physiology and pathophysiology of the brain, we employed chromis-1 to monitor cytoplasmic concentrations of labile ZnĀ­(II) in oligodendrocytes, an important cellular constituent of the brain, at different stages of development in cell culture. These studies revealed a decrease in probe saturation upon differentiation to mature oligodendrocytes, implying significant changes to cellular zinc homeostasis during maturation with an overall reduction in cellular zinc availability. Optimized for TPEM, chromis-1 is especially well-suited for exploring the role of labile zinc pools in live cells under a broad range of physiological and pathological conditions
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