An empirical quantitative fluorescence resonance energy transfer method for multiple acceptors based on partial acceptor photobleaching

被引:19
作者
Yu, Huaina [2 ,3 ]
Zhang, Jianwei [2 ,3 ]
Li, Huali [2 ,3 ]
Qu, Junle [1 ]
Chen, Tongsheng [2 ,3 ]
机构
[1] Shenzhen Univ, Inst Optoelect, Minist Educ & Guangdong Prov, Key Lab Optoelect Devices & Syst, Shenzhen 518060, Peoples R China
[2] S China Normal Univ, Coll Biophoton, MOE Key Lab Laser Life Sci, Guangzhou 510631, Guangdong, Peoples R China
[3] S China Normal Univ, Coll Biophoton, Inst Laser Life Sci, Guangzhou 510631, Guangdong, Peoples R China
基金
中国国家自然科学基金;
关键词
CASPASE-3; ACTIVATION; FRET; CELLS; YFP;
D O I
10.1063/1.4729481
中图分类号
O59 [应用物理学];
学科分类号
摘要
We present an empirical quantitative fluorescence resonance energy transfer (FRET) method based on partial acceptor photobleaching, termed emp-PbFRET. This method can be used to determine the FRET efficiency of construct with multiple acceptors by measuring the degree of acceptor photobleaching (x) and the donor intensity before and after photobleaching. We validated the emp-PbFRET method using the constructs consisting one Cerulean and two (VCV) or three (VCVV) Venus. The FRET efficiencies of VCV and VCVV in living cells obtained by emp-PbFRET method under different x (10%-60%) were consistent with those obtained by fluorescence lifetime imaging and spectral imaging methods. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.4729481]
引用
收藏
页数:4
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