Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

Alena Rudkouskaya; Nattawut Sinsuebphon; Xavier Intes; Joseph E. Mazurkiewicz; Margarida Barroso

doi:10.1117/12.2258231

21 February 2017 Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

Alena Rudkouskaya, Nattawut Sinsuebphon, Xavier Intes, Joseph E. Mazurkiewicz, Margarida Barroso

Proceedings Volume 10069, Multiphoton Microscopy in the Biomedical Sciences XVII; 1006917 (2017) https://doi.org/10.1117/12.2258231
Event: SPIE BiOS, 2017, San Francisco, California, United States

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.

Conference Presentation

Citation Download Citation

Alena Rudkouskaya, Nattawut Sinsuebphon, Xavier Intes, Joseph E. Mazurkiewicz, and Margarida Barroso "Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006917 (21 February 2017); https://doi.org/10.1117/12.2258231

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available