Simulation of the measurement error of independent emission-spectral unmixing FRET

Tingting Lin; Shuzhen Tang; Haoran Zhong; Yating Lin; Juqiang Lin; Yimei Huang

doi:10.1117/12.2573142

10 October 2020 Simulation of the measurement error of independent emission-spectral unmixing FRET

Tingting Lin, Shuzhen Tang, Haoran Zhong, Yating Lin, Juqiang Lin, Yimei Huang

Proceedings Volume 11553, Optics in Health Care and Biomedical Optics X; 115531L (2020) https://doi.org/10.1117/12.2573142
Event: SPIE/COS Photonics Asia, 2020, Online Only

Conference Poster

Abstract

Independent emission-spectral unmixing fluorescence resonance energy transfer, Iem-spFRET, is a novel and powerful tool for measuring FRET efficiency in real time. In this paper, we simulate the measurement error of the Iem-spFRET by introducing random noise in sample data, donor fingerprint, and acceptor fingerprint. The random noise intensity is set from 0.0005 to 0.0025, corresponding to 5% - 25% of the maximum donor fingerprint intensity. The simulated results show the effect of random noise on apparent FRET efficiency (E_fD) is less than on receptor-to-donor concentration ratio (R_c). Random noise with 10% maximum donor fingerprint intensity only leads to 0.33% variation of when the noise is added to both sample and fingerprints. These results indicate that Iem-spFRET is a robust method and could be applied on cases with weak FRET signal.

Citation Download Citation

Tingting Lin, Shuzhen Tang, Haoran Zhong, Yating Lin, Juqiang Lin, and Yimei Huang "Simulation of the measurement error of independent emission-spectral unmixing FRET", Proc. SPIE 11553, Optics in Health Care and Biomedical Optics X, 115531L (10 October 2020); https://doi.org/10.1117/12.2573142

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