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
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 donorāacceptor
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
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 donorāacceptor
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