Gate-width impact on NIR FRET lifetime fitting using gated ICCD

Sez-Jade Chen; Xavier Intes

doi:10.1117/12.2217780

10 March 2016 Gate-width impact on NIR FRET lifetime fitting using gated ICCD

Sez-Jade Chen, Xavier Intes

Proceedings Volume 9701, Multimodal Biomedical Imaging XI; 970117 (2016) https://doi.org/10.1117/12.2217780
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Förster Resonance Energy Transfer (FRET) is widely used to sense molecular interactions occurring at the nanoscale. In vitro and ex vivo protocols for visualizing FRET are already well-established, but in vivo studies have proven to be more challenging. One issue that hinders in vivo visualization of FRET is the higher absorption and scattering of visible light within tissues. In this case, light in the near-infrared (NIR) spectral window is required for increased depth sensing. Moreover, due to spectral variation in optical properties as well as heterogeneous spatial distribution, lifetime-based FRET imaging is preferred. Herein, we investigate the effect of temporal acquisition settings on the lifetime-based estimation of the fraction of quenched donor molecules (A1) as well as the quenched donor lifetime (τ₁). We performed in silico, in vitro, and in vivo experiments under gate widths of 300ps to 1000ps in 100ps intervals to determine the effect on quantification of A₁ and τ₁. Even though the NIR fluorescent dyes have shorter lifetimes then visible fluorophores, we were still able to accurately quantify FRET under all tested system gate widths and experimental conditions.

Citation Download Citation

Sez-Jade Chen and Xavier Intes "Gate-width impact on NIR FRET lifetime fitting using gated ICCD", Proc. SPIE 9701, Multimodal Biomedical Imaging XI, 970117 (10 March 2016); https://doi.org/10.1117/12.2217780

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KEYWORDS

Fluorescence resonance energy transfer

In vivo imaging

Near infrared

Photons

Picosecond phenomena

In vitro testing

Tumors

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