Stimulated Emission Properties of Fluorophores by CW-STED Single Molecule Spectroscopy

被引：16
作者：
Bouzin, Margaux [1 ]
Chirico, Giuseppe [1 ]
D'Alfonso, Laura [1 ]
Sironi, Laura [1 ]
Soavi, Giancarlo [2 ]
Cerullo, Giulio [2 ]
Campanini, Barbara [3 ]
Collini, Maddalena [1 ]
机构：
[1] Univ Milano Bicocca, Dept Phys, I-20126 Milan, Italy
[2] Politecn Milan, Dept Phys, IFN CNR, I-20133 Milan, Italy
[3] Univ Parma, Dept Pharmacol, I-43124 Parma, Italy
来源：
JOURNAL OF PHYSICAL CHEMISTRY B | 2013年 / 117卷 / 51期
关键词：
FLUORESCENCE MICROSCOPY; RESOLUTION; DEPLETION; DYNAMICS; STRATEGIES; EXCITATION; KINETICS;
D O I：
10.1021/jp409330t
中图分类号：
O64 [物理化学（理论化学）、化学物理学];
学科分类号：
070304 ; 081704 ;
摘要：
Fluorophores useful for STimulated Emission Depletion (STED) spectroscopy must fulfill strict requirements on depletion efficiency and photostability. These parameters determine the effective resolution of STED imaging. Resolution is typically measured on 30-80 nm spheres heavily decorated with STED bright fluorophores, limiting the possibility to estimate the true resolution achievable on a specific dye. Here we show how single molecule STED microscopy provides an estimate of the fluorophore stimulated emission cross section and of its photostability under STED irradiation. Fluorescein, a green and a yellow mutant of GFP, are tested, and the results are discussed and compared to those obtained with Chromeo488-covered 80 nm spheres on a commercial continuous-wave STED microscope.
引用
收藏
页码：16405 / 16415
页数：11
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