Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 54, Issue 50, Pages 15213-15217Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201507939
Keywords
fluorescent probes; phosphorus heterocycles; photophysics; photoresistance; STED microscopy
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Funding
- JST, CREST
- Japan Advanced Plant Science Network
- World Premier International Research Center (WPI) Initiative, Japan
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The development of stimulated emission depletion (STED) microscopy represented a major breakthrough in cellular and molecular biology. However, the intense laser beams required for both excitation and STED usually provoke rapid photobleaching of fluorescent molecular probes, which significantly limits the performance and practical utility of STED microscopy. We herein developed a photoresistant fluorescent dye C-Naphox as a practical tool for STED imaging. With excitation using either a lambda = 405 or 488 nm laser in protic solvents, C-Naphox exhibited an intense red/orange fluorescence (quantum yield phi(F) > 0.7) with a large Stokes shift (circa 5900 cm(-1)). Even after irradiation with a Xe lamp (300 W, lambda(ex) = 460 nm, full width at half maximum (FWHM) = 11 nm) for 12 hours, 99.5 % of C-Naphox remained intact. The high photoresistance of C-Naphox allowed repeated STED imaging of HeLa cells. Even after recording 50 STED images, 83 % of the initial fluorescence intensity persisted.
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