2. Academic Validation
  3. Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy

Bio-orthogonal Red and Far-Red Fluorogenic Probes for Wash-Free Live-Cell and Super-resolution Microscopy

  • ACS Cent Sci. 2021 Sep 22;7(9):1561-1571. doi: 10.1021/acscentsci.1c00703.
Philipp Werther 1,  Klaus Yserentant 2 3 4 5,  Felix Braun 2,  Kristin Grußmayer 6 7,  Vytautas Navikas 6,  Miao Yu 8 9 10,  Zhibin Zhang 2 11,  Michael J Ziegler 1 12,  Christoph Mayer 1,  Antoni J Gralak 1,  Marvin Busch 1,  Weijie Chi 13,  Frank Rominger 14,  Aleksandra Radenovic 6,  Xiaogang Liu 13,  Edward A Lemke 8 9 10,  Tiago Buckup 2,  Dirk-Peter Herten 2 3 4,  Richard Wombacher 1 12
Affiliations

Affiliations

  • 1 Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany.
  • 2 Institute of Physical Chemistry, Heidelberg University, Im Neuenheimer Feld 229, 69120 Heidelberg, Germany.
  • 3 Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Edgbaston, B15 2TT Birmingham, U.K.
  • 4 College of Medical and Dental Sciences, Medical School & School of Chemistry, University of Birmingham, Edgbaston, B15 2TT Birmingham, U.K.
  • 5 Faculty of Biosciences, Heidelberg University, Im Neuenheimer Feld 234, 69120 Heidelberg, Germany.
  • 6 Laboratory of Nanoscale Biology, École Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
  • 7 Delft University of Technology, Kavli Institute of Nanoscience, Department of Bionanoscience, Van der Maasweg 9, 2629 HZ Delft, The Netherlands.
  • 8 Biocenter Mainz, Departments of Biology and Chemistry, Johannes Gutenberg University, Hanns-Dieter-Hüsch-Weg 17, 55128 Mainz, Germany.
  • 9 Institute of Molecular Biology, Ackermannweg 4, 55128 Mainz, Germany.
  • 10 Structural and Computational Biology Unit, Cell Biology and Biophysics Unit, EMBL, Meyerhofstraße 1, 69117 Heidelberg, Germany.
  • 11 National Key Laboratory of Science and Technology on Tunable Laser, Harbin Institute of Technology, Building 2A, Yikuang Str.2, Harbin 150080, China.
  • 12 Department of Chemical Biology, Max Planck Institute for Medical Research, Jahnstraße 29, 69120 Heidelberg, Germany.
  • 13 Fluorescence Research Group, Singapore University of Technology and Design, 8 Somapah Road, 487372 Singapore.
  • 14 Institute of Organic Chemistry, Heidelberg University, Im Neuenheimer Feld 270, 69120 Heidelberg, Germany.
PMID: 34584958 DOI: 10.1021/acscentsci.1c00703
Abstract

Small-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labeling via bio-orthogonal tetrazine chemistry combines minimal label size with rapid labeling kinetics. At the same time, fluorogenic tetrazine-dye conjugates exhibit efficient quenching of dyes prior to target binding. However, live-cell compatible long-wavelength fluorophores with strong fluorogenicity have been difficult to realize. Here, we report close proximity tetrazine-dye conjugates with minimal distance between tetrazine and the fluorophore. Two synthetic routes give access to a series of cell-permeable and -impermeable dyes including highly fluorogenic far-red emitting derivatives with electron exchange as the dominant excited-state quenching mechanism. We demonstrate their potential for live-cell imaging in combination with unnatural Amino acids, wash-free multicolor and super-resolution STED, and SOFI imaging. These dyes pave the way for advanced fluorescence imaging of biomolecules with minimal label size.

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