@article{ab70bde506814cb0af8e162935a8f8a3,
title = "Photoactivatable Fluorophore for Stimulated Emission Depletion (STED) Microscopy and Bioconjugation Technique for Hydrophobic Labels",
abstract = "The use of photoactivatable dyes in STED microscopy has so far been limited by two-photon activation through the STED beam and by the fact that photoactivatable dyes are poorly solvable in water. Herein, we report ONB-2SiR, a fluorophore that can be both photoactivated in the UV and specifically de-excited by STED at 775 nm. Likewise, we introduce a conjugation and purification protocol to effectively label primary and secondary antibodies with moderately water-soluble dyes. Greatly reducing dye aggregation, our technique provides a defined and tunable degree of labeling, and improves the imaging performance of dye conjugates in general.",
keywords = "fluorescence, optical superresolution, photoactivation, protein labelling, stimulated emission depletion (STED)",
author = "Michael Weber and Khan, \{Taukeer A.\} and Patalag, \{Lukas J.\} and Mariano Bossi and Marcel Leutenegger and Belov, \{Vladimir N.\} and Hell, \{Stefan W.\}",
note = "Funding Information: The authors acknowledge the financial support provided by the German Bundesministerium f{\"u}r Bildung und Forschung (grant FKZ 13N14122). We thank Dr. Alexey N. Butkevich for fruitful discussions and starting compounds, and Dr. Mark Bates for providing the microscope and support for the PALM measurements. We thank Jens Schimpfhauser and Jan Seikowski (Chemistry Facility, MPI BPC) for the synthesis of intermediates. We are indebted to J{\"u}rgen Bienert (Chemistry Facility, MPI BPC), Dr. H. Frauendorf, Dr. M. John and co‐workers (Institut f{\"u}r organische und biomolekulare Chemie, University of G{\"o}ttingen, Germany) for recording spectra. We thank Dr. Stefan Stoldt for help with the antibody selection and Dr. Steffen J. Sahl for advice on the manuscript. We thank Dr. Ellen Rothermel and Tanja Gilat for assistance with cell culture and the preparation of labeled cells. Open access funding enabled and organized by Projekt DEAL. Funding Information: The authors acknowledge the financial support provided by the German Bundesministerium f?r Bildung und Forschung (grant FKZ 13N14122). We thank Dr. Alexey N. Butkevich for fruitful discussions and starting compounds, and Dr. Mark Bates for providing the microscope and support for the PALM measurements. We thank Jens Schimpfhauser and Jan Seikowski (Chemistry Facility, MPI BPC) for the synthesis of intermediates. We are indebted to J?rgen Bienert (Chemistry Facility, MPI BPC), Dr. H. Frauendorf, Dr. M. John and co-workers (Institut f?r organische und biomolekulare Chemie, University of G?ttingen, Germany) for recording spectra. We thank Dr. Stefan Stoldt for help with the antibody selection and Dr. Steffen J. Sahl for advice on the manuscript. We thank Dr. Ellen Rothermel and Tanja Gilat for assistance with cell culture and the preparation of labeled cells. Open access funding enabled and organized by Projekt DEAL. Publisher Copyright: {\textcopyright} 2020 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH",
year = "2021",
month = jan,
day = "4",
doi = "10.1002/chem.202004645",
language = "English",
volume = "27",
pages = "451--458",
journal = "Chemistry - A European Journal",
issn = "0947-6539",
publisher = "Wiley-VCH Verlag GmbH \& Co. KGaA",
number = "1",
}