Determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells

Zexian Hou; Yuhua Wang; Liqin Zheng; Tongsheng Chen; Hongqin Yang; Shusen Xie

doi:10.1117/12.2245543

31 October 2016 Determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells

Zexian Hou, Yuhua Wang, Liqin Zheng, Tongsheng Chen, Hongqin Yang, Shusen Xie

Author Affiliations +

Proceedings Volume 10024, Optics in Health Care and Biomedical Optics VII; 1002428 (2016) https://doi.org/10.1117/12.2245543
Event: SPIE/COS Photonics Asia, 2016, Beijing, China

Abstract

The cross section is a significant parameter for fluorescence protein and determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells is the vital issue for two-photon excitation FRET quantification. In this study, Hela cells were labeled with FPs that acceptor-to-donor concentration ratio is 1 to 1 and acceptor-to-donor cross section ratio ranged from 700nm to 960nm was obtained by emission spectral unmixing with independent excitation crosstalk correction. The results show that acceptor-to-donor cross section ratio declines with the excitation wavelength from 700nm to 790nm and then increases inversely from 790nm to 960nm. This method can quickly determine the cross section without any additional references, which could provide a powerful and convenient tool for measuring acceptor-to-donor cross section ratio by two-photon excitation in living cells.

Citation Download Citation

Zexian Hou, Yuhua Wang, Liqin Zheng, Tongsheng Chen, Hongqin Yang, and Shusen Xie "Determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells", Proc. SPIE 10024, Optics in Health Care and Biomedical Optics VII, 1002428 (31 October 2016); https://doi.org/10.1117/12.2245543

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