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Time-gated Fluorescence Lifetime Imaging in the Near Infrared Regime; A Comprehensive Study Toward In Vivo Imaging

Meital Harel, Uri Arbiv, View ORCID ProfileRinat Ankri
doi: https://doi.org/10.1101/2023.05.21.541614
Meital Harel
1Department of Physics, Ariel University, Ariel, Israel
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Uri Arbiv
1Department of Physics, Ariel University, Ariel, Israel
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Rinat Ankri
1Department of Physics, Ariel University, Ariel, Israel
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  • ORCID record for Rinat Ankri
  • For correspondence: rinatsel@ariel.ac.il
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Abstract

Fluorescence lifetime imaging has an enormous impact on our understanding of biological systems, both in vitro and in vivo. It is a powerful tool for the non-invasive in vitro and in vivo biomolecular and cellular investigations. In particular, it has the potential to target and multiplex different species with high sensitivity and specificity, providing a fast and noninvasive readout at low cost. In this work, we present a time-saving Monte Carlo (MC) simulation of fluorescent photons scattering within a turbid medium, followed by phasor analyzes which enabled the simple multiplexing of different targets in one frame. We then demonstrate a simple and fast method for wide-field FLI in the near-infrared (NIR) region, where tissue scattering and autofluorescence are significantly lower, to enable imaging of deep tissue, using the state-of-the-art timed single-photon avalanche diode array camera (SPAD), SPAD512S. In particular, we show how phasor scattering increases with depth. However, using appropriate background correction, a simple “cut-off” method, and averaging, we can multiplex two targets in one image to a depth of 1 cm in tissue. Our results show that it is possible to perform in vivo FLI under challenging conditions, using standard NIR fluorophores with short lifetimes.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 22, 2023.
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Time-gated Fluorescence Lifetime Imaging in the Near Infrared Regime; A Comprehensive Study Toward In Vivo Imaging
Meital Harel, Uri Arbiv, Rinat Ankri
bioRxiv 2023.05.21.541614; doi: https://doi.org/10.1101/2023.05.21.541614
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Time-gated Fluorescence Lifetime Imaging in the Near Infrared Regime; A Comprehensive Study Toward In Vivo Imaging
Meital Harel, Uri Arbiv, Rinat Ankri
bioRxiv 2023.05.21.541614; doi: https://doi.org/10.1101/2023.05.21.541614

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