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Proceedings Paper

Comparison of NIR FRET pairs for quantitative transferrin-based assay
Author(s): Nattawut Sinsuebphon; Travis Bevington; Lingling Zhao; Abe Ken; Margarida Barroso; Xavier Intes
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Paper Abstract

Transferrin (Tfn) is commonly used as a drug delivery carrier for cancer treatment. Tfn cellular internalization can be observed by Förster resonance energy transfer (FRET), which occurs when two fluorophores - donor and acceptor - are a few nanometers apart. Donor fluorescence lifetime can be used to sense and quantify FRET occurrence. In FRET state, the donor is quenched leading to a significant reduction in its lifetime. In this study, donor and acceptor near-infrared (NIR) fluorophore-labeled Tfn were used to quantify cellular internalization in breast cancer cell line (T47D). Based on donor lifetime, quantum yield and spectral data, seven NIR FRET pairs were chosen for this comparison. Performance of the different NIR FRET pairs was evaluated in vitro in multiwell plate settings and by analyzing the relationship between quenched donor fraction and acceptor:donor ratio. Additionally, we performed brightness comparison between each pairs. Several parameters, such as brightness, lifetime, R0 and FRET donor population values are used to identify the most suitable NIR FRET pair for in vivo studies in preclinical settings.

Paper Details

Date Published: 17 February 2014
PDF: 7 pages
Proc. SPIE 8937, Multimodal Biomedical Imaging IX, 89370X (17 February 2014); doi: 10.1117/12.2040097
Show Author Affiliations
Nattawut Sinsuebphon, Rensselaer Polytechnic Institute (United States)
Travis Bevington, Albany Medical College (United States)
Lingling Zhao, Rensselaer Polytechnic Institute (United States)
Abe Ken, Albany Medical College (United States)
Margarida Barroso, Albany Medical College (United States)
Xavier Intes, Rensselaer Polytechnic Institute (United States)

Published in SPIE Proceedings Vol. 8937:
Multimodal Biomedical Imaging IX
Fred S. Azar; Xavier Intes, Editor(s)

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