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Effects of donor and acceptor's fluorescence lifetimes on the method of applying Förster resonance energy transfer in STED microscopy
Author(s) -
DENG SUHUI,
CHEN JIANFANG,
GAO ZHAOSHUAI,
FAN CHUNHAI,
YAN QIURONG,
WANG YUHAO
Publication year - 2018
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12608
Subject(s) - sted microscopy , förster resonance energy transfer , microscopy , energy transfer , fluorescence , acceptor , fluorescence microscope , chemistry , photochemistry , molecular physics , stimulated emission , optics , physics , laser , condensed matter physics
Summary Förster resonance energy transfer (FRET) probes being used to improve the resolution of stimulated emission depletion (STED) microscopy are numerically discussed. Besides the FRET efficiency and the excitation intensity, the fluorescence lifetimes of donor and acceptor are found to be another key parameter for the resolution enhancement. Using samples of FRET pairs with shorter donor lifetime and longer acceptor lifetime enhances the nonlinearity of the donor fluorescence, which leads to an increased resolution. The numerical simulation shows that a double resolution improvement of STED microscopy can be achieved by using Cy3–Atto647N samples when compared with that of using standard Cy3‐only samples.