28 August 2014 Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer
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Abstract
Fluorescence lifetime imaging (FLIM) aims at quantifying the exponential decay rate of fluorophores to yield lifetime maps over the imaged sample. When combined with Förster resonance energy transfer (FRET), the technique can be used to indirectly sense interactions at the nanoscale such as protein–protein interactions, protein–DNA interactions, and protein conformational changes. In the case of FLIM-FRET, the fluorescence intensity decays are fitted to a biexponential model in order to estimate the lifetime and fractional amplitude coefficients of each component of the population of the donor fluorophore (quenched and nonquenched). Numerous time data points, also called temporal or time gates, are typically employed for accurately estimating the model parameters, leading to lengthy acquisition times and significant computational demands. This work investigates the effect of the number and location of time gates on model parameter estimation accuracy. A detailed model of a FLIM-FRET imaging system is used for the investigation, and the simulation outcomes are validated with
© 2014 Society of Photo-Optical Instrumentation Engineers (SPIE)
Travis Omer, Travis Omer, Lingling Zhao, Lingling Zhao, Xavier Intes, Xavier Intes, Juergen Hahn, Juergen Hahn, } "Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer," Journal of Biomedical Optics 19(8), 086023 (28 August 2014). https://doi.org/10.1117/1.JBO.19.8.086023 . Submission:
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