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Journal of Biomedical Optics

Spectral wide-field microscopic fluorescence resonance energy transfer imaging in live cells
Author(s): Lili Zhang; Guiqi Qin; Liuying Chai; Jiang Zhang; Fangfang Yang; Hongqin Yang; Shusen Xie; Tongsheng Chen
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Paper Abstract

With its precise, sensitive, and nondestructive features, spectral unmixing-based fluorescence resonance energy transfer (FRET) microscopy has been widely applied to visualize intracellular biological events. In this report, we set up a spectral wide-field microscopic FRET imaging system by integrating a varispec liquid crystal tunable filter into a wide-field microscope for quantitative FRET measurement in living cells. We implemented a representative emission-spectral unmixing-based FRET measurement method on this platform to simultaneously acquire pixel-to-pixel images of both FRET efficiency (E) and acceptor-to-donor concentration ratio (RC) in living HepG2 cells expressing fusion proteins in the presence or absence of free donors and acceptors and obtained consistent results with other instruments and methods. This stable and low-cost spectral wide-field microscopic FRET imaging system provides a new toolbox for imaging molecular events with high spatial resolution in living cells.

Paper Details

Date Published: 17 August 2015
PDF: 9 pages
J. Biomed. Opt. 20(8) 086011 doi: 10.1117/1.JBO.20.8.086011
Published in: Journal of Biomedical Optics Volume 20, Issue 8
Show Author Affiliations
Lili Zhang, South China Normal Univ. (China)
Guiqi Qin, South China Normal Univ. (China)
Liuying Chai, South China Normal Univ. (China)
Jiang Zhang, South China Normal University (China)
Fangfang Yang, South China Normal Univ. (China)
Hongqin Yang, Fujian Normal University (China)
Shusen Xie, Fujian Normal Univ. (China)
Tongsheng Chen, South China Normal Univ. (China)

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