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Proceedings Paper

Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo
Author(s): Alena Rudkouskaya; Nattawut Sinsuebphon; Xavier Intes; Joseph E. Mazurkiewicz; Margarida Barroso
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Paper Abstract

To guide the development of targeted therapies with improved efficacy and accelerated clinical acceptance, novel imaging methodologies need to be established. Toward this goal, fluorescence lifetime Förster resonance energy transfer (FLIM-FRET) imaging assays capitalize on the ability of antibodies or protein ligands to bind dimerized membrane bound receptors to measure their target engagement levels in cancer cells. Conventional FLIM FRET microscopy has been widely applied at visible wavelengths to detect protein-protein interactions in vitro. However, operation at these wavelengths restricts imaging quality and ability to quantitate lifetime changes in in vivo small animal optical imaging due to high auto-fluorescence and light scattering. Here, we have analyzed the uptake of iron-bound transferrin (Tf) probes into human breast cancer cells using FLIM-FRET microscopy in the visible and near-infrared (NIR) range. The development of NIR FLIM FRET microscopy allows for the use of quantitative lifetime-based molecular assays to measure drug-target engagement levels at multiple scales: from in vitro microscopy to in vivo small animal optical imaging (macroscopy). This novel approach can be extended to other receptors, currently targeted in oncology. Hence, lifetime-based molecular imaging can find numerous applications in drug delivery and targeted therapy assessment and optimization.

Paper Details

Date Published: 21 February 2017
PDF: 8 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006917 (21 February 2017); doi: 10.1117/12.2258231
Show Author Affiliations
Alena Rudkouskaya, Albany Medical College (United States)
Nattawut Sinsuebphon, Rensselaer Polytechnic Institute (United States)
Xavier Intes, Rensselaer Polytechnic Institute (United States)
Joseph E. Mazurkiewicz, Albany Medical College (United States)
Margarida Barroso, Albany Medical College (United States)


Published in SPIE Proceedings Vol. 10069:
Multiphoton Microscopy in the Biomedical Sciences XVII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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