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

Junle Qu; Ziyang Lin; Lixin Liu; Xuan Guo; Danni Chen; Hanben Niu

doi:10.1117/12.569582

18 January 2005 Two-photon excited fluorescence lifetime imaging microscopy for FRET study on protein interactions

Junle Qu, Ziyang Lin, Lixin Liu, Xuan Guo, Danni Chen, Hanben Niu

Author Affiliations +

Proceedings Volume 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II; (2005) https://doi.org/10.1117/12.569582
Event: Photonics Asia, 2004, Beijing, China

Abstract

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 & Hickl 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.

Citation Download Citation

Junle Qu, Ziyang Lin, Lixin Liu, Xuan Guo, Danni Chen, and Hanben Niu "Two-photon excited fluorescence lifetime imaging microscopy for FRET study on protein interactions", Proc. SPIE 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II, (18 January 2005); https://doi.org/10.1117/12.569582

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