Fluorescent rhodamines and fluorogenic carbopyronines for super-resolution STED microscopy in living cells Journal Article


Author(s): Butkevich, Alexey N; Mitronova, Gyuzel Y; Sidenstein, Sven C; Klocke, Jessica L; Kamin, Dirk; Meineke, Dirk N; D'Este, Elisa; Kraemer, Philip T; Danzl, Johannes G; Belov, Vladimir N; Hell, Stefan W
Article Title: Fluorescent rhodamines and fluorogenic carbopyronines for super-resolution STED microscopy in living cells
Affiliation
Abstract: A range of bright and photostable rhodamines and carbopyronines with absorption maxima in the range of λ=500-630 nm were prepared, and enabled the specific labeling of cytoskeletal filaments using HaloTag technology followed by staining with 1 μm solutions of the dye-ligand conjugates. The synthesis, photophysical parameters, fluorogenic behavior, and structure-property relationships of the new dyes are discussed. Light microscopy with stimulated emission depletion (STED) provided one- and two-color images of living cells with an optical resolution of 40-60 nm.
Keywords: Fluorescence; carbopyronines; living cells; optical microscopy; rhodamines
Journal Title: Angewandte Chemie - International Edition
Volume: 55
Issue 10
ISSN: 1521-3773
Publisher: Wiley-Blackwell  
Date Published: 2016-03-01
Start Page: 3290
End Page: 3294
Copyright Statement: CC BY-NC-ND 4.0
DOI: 10.1002/anie.201511018
Notes: We thank Prof. Y. Okada (RIKEN Quantitative Biology Center, Osaka, Japan) for the gift of β-tubulin-Halo plasmid, T. Gilat and Dr. E. Rothermel (MPIBPC, Göttingen, Germany) for cell culture and transfection, M. Pulst, J. Bienert (MPIBPC), Dr. M. John, Dr. H. Frauendorf, and co-workers (Institut für Organische und Biomolekulare Chemie, Georg-August-Universität, Göttingen, Germany) for UV/Vis, NMR, and ESI-MS spectra, Prof. M. L. Bossi (University of Buenos-Aires, Argentina) for measuring fluorescence lifetimes, and Dr. S. Vos and Prof. P. Cramer (MPIBPC) for access to a Tecan microplate reader. S.W.H. acknowledges a grant from the Bundesministerium für Bildung und Forschung (BMBF 513) within the program “Optische Technologien für Biowissenschaften und Gesundheit” (FKZ 13N11066). J.G.D. was supported by funds from the People Programme (Marie Curie Actions) of the European Union's Seventh Framework Programme (FP7/2007–2013; REA grant agreement PIEF-GA-2011-299283).
Open access: yes (OA journal)