Real‐time fiber‐based fluorescence lifetime imaging with synchronous external illumination: A new path for clinical translation

Year: 2020

Authors: Lagarto Joao L.; Shcheslavskiy Vladislav; Pavone Francesco S.; Cicchi Riccardo

Autors Affiliation: Natl Res Council INO CNR, Natl Inst Opt, Via Nello Carrara 1, I-50019 Sesto Fiorentino, Italy; European Lab Nonlinear Spect Lens, Sesto Fiorentino, Italy; Becker & Hickl GmbH, Berlin, Germany; Univ Florence, Dept Phys, Sesto Fiorentino, Italy.

Abstract: Time-correlated single photon counting is the “gold-standard” method for fluorescence lifetime measurements and has demonstrated potential for clinical deployment. However, the translation of the technology into clinic is hindered by the use of ultrasensitive detectors, which make the fluorescence acquisition impractical with bright lighting conditions such as in clinical settings. We address this limitation by interleaving periodic fluorescence detection with synchronous out-of-phase externally modulated light source, thus guaranteeing specimen illumination and a fluorescence signal free from bright background light upon temporal separation. Fluorescence lifetime maps are generated in real-time from single-point measurements by tracking a reference beam and using the phasor approach. We demonstrate the feasibility and practicality of this technique in a number of biological
specimens, including real-time mapping of degraded articular cartilage. This method is compatible and can be integrated with existing clinical microscopic, endoscopic and robotic modalities, thus offering a new pathway towards label-free diagnostics and surgical guidance in a number of clinical applications.

Journal/Review: JOURNAL OF BIOPHOTONICS

Volume: 13 (3)      Pages from: e201960119-1  to: e201960119-14

More Information: H2020, Grant/Award Numbers: 654148 Laserlab-Europe, H2020-ICT-2016-1 732111 PICCOLO; Regione Toscana, Grant/Award Numbers: PAR FAS 2007-2013 -Bando FAS Salute 2014, POR FSE 2014-2020 Giovanisi
KeyWords: fiber optics, fluorescence lifetime imaging, phasors, surgical guidance, TCSPC, tissue
spectroscopy
DOI: 10.1002/jbio.201960119

Citations: 16
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