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Hauke S, von Appen A, Quidwai T, et al. Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells. Chem Sci. 2016;8(1):559-566doi: 10.1039/c6sc02088g.
Hauke, S., von Appen, A., Quidwai, T., Ries, J., & Wombacher, R. (2017). Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells. Chemical science, 8(1), 559-566. https://doi.org/10.1039/c6sc02088g
Hauke, Sebastian, et al. "Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells." Chemical science vol. 8,1 (2017): 559-566. doi: https://doi.org/10.1039/c6sc02088g
Hauke S, von Appen A, Quidwai T, Ries J, Wombacher R. Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells. Chem Sci. 2017 Jan 01;8(1):559-566. doi: 10.1039/c6sc02088g. Epub 2016 Sep 05. PMID: 28451202; PMCID: PMC5351804.
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Chem Sci. 2017 Jan 01;8(1):559-566. doi: 10.1039/c6sc02088g. Epub 2016 Sep 05.

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

Chemical science

Sebastian Hauke, Alexander von Appen, Tooba Quidwai, Jonas Ries, Richard Wombacher

Affiliations

  1. Institute of Pharmacy and Molecular Biotechnology , Ruprecht-Karls-University Heidelberg , Im Neuenheimer Feld 364 , 69120 Heidelberg , Germany . Email: [email protected] ; ; Tel: +49 6221 544879.
  2. European Molecular Biology Laboratory , Meyerhofstraße 1 , 69117 Heidelberg , Germany.

PMID: 28451202 PMCID: PMC5351804 DOI: 10.1039/c6sc02088g

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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