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Journal of Biomedical Optics • Open Access

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer
Author(s): Travis Omer; Lingling Zhao; Xavier Intes; Juergen Hahn

Paper 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 in vitro and in vivo experimental data. In all cases investigated, it is found that 10 equally spaced time gates allow robust estimation of model-based parameters with accuracy similar to that of full temporal datasets (90 gates).

Paper Details

Date Published: 28 August 2014
PDF: 9 pages
J. Biomed. Opt. 19(8) 086023 doi: 10.1117/1.JBO.19.8.086023
Published in: Journal of Biomedical Optics Volume 19, Issue 8
Show Author Affiliations
Travis Omer, Rensselaer Polytechnic Institute (United States)
Lingling Zhao, Rensselaer Polytechnic Institute (United States)
Xavier Intes, Rensselaer Polytechnic Institute (United States)
Juergen Hahn, Rensselaer Polytechnic Institute (United States)

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