Two-photon excited fluorescence lifetime imaging microscopy for FRET study on protein interactions

被引:1
|
作者
Qu, JL [1 ]
Lin, ZY [1 ]
Liu, LX [1 ]
Guo, X [1 ]
Chen, DN [1 ]
Niu, HB [1 ]
机构
[1] Shenzhen Univ, Inst Optoelect, Shenzhen 518060, Peoples R China
关键词
fluorescence lifetime imaging; fluorescence resonance energy transfer; time-correlated single photon counting; two-photon excited fluorescence;
D O I
10.1117/12.569582
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Two-photon excited fluorescence lifetime imaging (2P-FLIM) provides a more direct and precise approach to fluorescence resonance energy transfer (FRET), which allows studying the dynamic behavior of protein-protein interactions in living cells. In this paper, we describe the combination of a Leica TCS SP2 laser scanning microscope and a time-correlated single photon counting (TCSPC) lifetime imaging module developed by Becker & Hick] for two-photon excited fluorescence lifetime imaging. This 2P-FLIM system was used for FRET study on the interaction of heat shock protein hsp27 with p38 MAP kinase in the single living cell. Results show that the reduction in donor (CFP) lifetime in the presence of acceptor (YFP) reveals interactions between the two proteins.
引用
收藏
页码:517 / 522
页数:6
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