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High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria.

Pedretti, Ettore and Tanner, Michael G. and Choudhary, Tushar R. and Krstajić, Nikola and Megia-Fernandez, Alicia and Henderson, Robert K. and Bradley, Mark and Thomson, Robert R. and Girkin, John M. and Dhaliwal, Kevin and Dalgarno, Paul A. (2019) 'High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria.', Biomedical optics express., 10 (1). pp. 181-195.

Abstract

We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence imaging by temporally shifting the dual excitation lasers, with respect to each other, to separate the two spectrally distinct fluorescent decays in time. Combining the temporal encoding, to provide spectral separation, with lifetime measurements we show a one FPS, multi-channel endomicroscopy platform for clinical applications and diagnosis. We demonstrate the potential of the system by imaging SmartProbe labeled bacteria in ex vivo samples of human lung using lifetimeto differentiate bacterial fluorescence from the strong background lung autofluorescence which was used to provide structural information.

Item Type:Article
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Available under License - Creative Commons Attribution.
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Status:Peer-reviewed
Publisher Web site:https://doi.org/10.1364/BOE.10.000181
Publisher statement:Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.
Date accepted:24 November 2018
Date deposited:13 December 2018
Date of first online publication:12 December 2018
Date first made open access:No date available

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