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A Phosphole Oxide Based Fluorescent Dye with Exceptional Resistance to Photobleaching: A Practical Tool for Continuous Imaging in STED Microscopy
Author(s) -
Wang Chenguang,
Fukazawa Aiko,
Taki Masayasu,
Sato Yoshikatsu,
Higashiyama Tetsuya,
Yamaguchi Shigehiro
Publication year - 2015
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201507939
Subject(s) - sted microscopy , photobleaching , fluorescence , microscopy , stimulated emission , fluorescence microscope , chemistry , laser , fluorescence lifetime imaging microscopy , optics , materials science , physics
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 λ =405 or 488 nm laser in protic solvents, C‐Naphox exhibited an intense red/orange fluorescence (quantum yield Φ F >0.7) with a large Stokes shift (circa 5900 cm −1 ). Even after irradiation with a Xe lamp (300 W, λ 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.