AI Article Synopsis
- New fluorophore-conjugates were developed for dual-color photoactivation and super-resolution imaging in live mammalian cells, designed for specific localization to intracellular targets.
- Researchers employed precise protein labeling techniques to attach light-activated fluorophores, allowing for visualization of various proteins and structures within living cells using UV light.
- The probes enabled advanced techniques like inverse fluorescence recovery after photo-bleaching (iFRAP) and photo-activated localization microscopy (PALM), providing enhanced real-time insights into protein dynamics and improved localization precision.
Article Abstract
We present new fluorophore-conjugates for dual-color photoactivation and super-resolution imaging inside live mammalian cells. These custom-designed, photo-caged Q-rhodamines and fluoresceins are cell-permeable, bright and localize specifically to intracellular targets. We utilized established orthogonal protein labeling strategies to precisely attach the photoactivatable fluorophores to proteins with subsequent activation of fluorescence by irradiation with UV light. That way, diffusive cytosolic proteins, histone proteins as well as filigree mitochondrial networks and focal adhesion proteins were visualized inside living cells. We applied the new photoactivatable probes in inverse fluorescence recovery after photo-bleaching (iFRAP) experiments, gaining real-time access to protein dynamics from live biological settings with resolution in space and time. Finally, we used the caged Q-rhodamine for photo-activated localization microscopy (PALM) on both fixed and live mammalian cells, where the superior molecular brightness and photo-stability directly resulted in improved localization precisions for different protein targets.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5351804 | PMC |
| http://dx.doi.org/10.1039/c6sc02088g | DOI Listing |
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