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

Pedretti, Ettore; Tanner, Michael G.; Choudhary, Tushar R.; Krstajić, Nikola; Megia-Fernandez, Alicia; Henderson, Robert K.; Bradley, Mark; Thomson, Robert R.; Girkin, John M.; Dhaliwal, Kevin; Dalgarno, Paul A.

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


Authors

Ettore Pedretti

Michael G. Tanner

Tushar R. Choudhary

Nikola Krstajić

Alicia Megia-Fernandez

Robert K. Henderson

Mark Bradley

Robert R. Thomson

Kevin Dhaliwal

Paul A. Dalgarno



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.

Citation

Pedretti, E., Tanner, M. G., Choudhary, T. R., Krstajić, N., Megia-Fernandez, A., Henderson, R. K., …Dalgarno, P. 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), 181-195. https://doi.org/10.1364/boe.10.000181

Journal Article Type Article
Acceptance Date Nov 24, 2018
Online Publication Date Dec 12, 2018
Publication Date Jan 1, 2019
Deposit Date Dec 12, 2018
Publicly Available Date Dec 13, 2018
Journal Biomedical Optics Express
Publisher Optica
Peer Reviewed Peer Reviewed
Volume 10
Issue 1
Pages 181-195
DOI https://doi.org/10.1364/boe.10.000181

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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright 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.





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