In vitro and in vivo phasor analysis of stoichiometry and pharmacokinetics using short‐lifetime near‐infrared dyes and time‐gated imaging

Chen, Sez‐Jade; Sinsuebphon, Nattawut; Rudkouskaya, Alena; Barroso, Margarida; Intes, Xavier; Michalet, Xavier

doi:10.1002/jbio.201800185

Title: In vitro and in vivo phasor analysis of stoichiometry and pharmacokinetics using short‐lifetime near‐infrared dyes and time‐gated imaging

Journal Article · 09 December 2018 · Journal of Biophotonics

DOI: https://doi.org/10.1002/jbio.201800185 · OSTI ID:1485161

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Department of Biomedical Engineering Rensselaer Polytechnic Institute Troy New York
Department of Molecular and Cellular Physiology Albany Medical College Albany New York
Department of Chemistry and Biochemistry University of California at Los Angeles Los Angeles California

We introduce a simple new approach for time‐resolved multiplexed analysis of complex systems using near‐infrared (NIR) dyes, applicable to in vitro and in vivo studies. We show that fast and precise in vitro quantification of NIR fluorophores' short (subnanosecond) lifetime and stoichiometry can be done using phasor analysis, a computationally efficient and user‐friendly representation of complex fluorescence intensity decays obtained with pulsed laser excitation and time‐gated camera imaging. We apply this approach to the study of binding equilibria by Förster resonant energy transfer using two different model systems: primary/secondary antibody binding in vitro and ligand/receptor binding in cell cultures. We then extend it to dynamic imaging of the pharmacokinetics of transferrin engagement with the transferrin receptor in live mice, elucidating the kinetics of differential transferrin accumulation in specific organs, straightforwardly differentiating specific from nonspecific binding. Our method, implemented in a freely‐available software, has the advantage of time‐resolved NIR imaging, including better tissue penetration and background‐free imaging, but simplifies and considerably speeds up data processing and interpretation, while remaining quantitative. These advances make this method attractive and of broad applicability for in vitro and in vivo molecular imaging and could be extended to applications as diverse as image‐guided surgery or optical tomography.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: DE‐FC02‐02ER63421

OSTI ID:: 1485161

Journal Information:: Journal of Biophotonics, Journal Name: Journal of Biophotonics Vol. 12 Journal Issue: 3; ISSN 1864-063X

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: Germany

Language:: English

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