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Issue 1, 2017
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Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells

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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.

Graphical abstract: Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells

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Publication details

The article was received on 12 May 2016, accepted on 01 Sep 2016 and first published on 05 Sep 2016


Article type: Edge Article
DOI: 10.1039/C6SC02088G
Citation: Chem. Sci., 2017,8, 559-566
  • Open access: Creative Commons BY-NC license
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    Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells

    S. Hauke, A. von Appen, T. Quidwai, J. Ries and R. Wombacher, Chem. Sci., 2017, 8, 559
    DOI: 10.1039/C6SC02088G

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