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Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue

Krstajić, Nikola; Akram, Ahsan R.; Choudhary, Tushar R.; McDonald, Neil; Tanner, Michael G.; Pedretti, Ettore; Dalgarno, Paul A.; Scholefield, Emma; Girkin, John M.; Moore, Anne; Bradley, Mark; Dhaliwal, Kevin

Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue Thumbnail


Authors

Nikola Krstajić

Ahsan R. Akram

Tushar R. Choudhary

Neil McDonald

Michael G. Tanner

Ettore Pedretti

Paul A. Dalgarno

Emma Scholefield

Anne Moore

Mark Bradley

Kevin Dhaliwal



Abstract

We demonstrate a fast two-color widefield fluorescence microendoscopy system capable of simultaneously detecting several disease targets in intact human ex vivo lung tissue. We characterize the system for light throughput from the excitation light emitting diodes, fluorescence collection efficiency, and chromatic focal shifts. We demonstrate the effectiveness of the instrument by imaging bacteria (Pseudomonas aeruginosa) in ex vivo human lung tissue. We describe a mechanism of bacterial detection through the fiber bundle that uses blinking effects of bacteria as they move in front of the fiber core providing detection of objects smaller than the fiber core and cladding (∼3  μm ∼3  μm ). This effectively increases the measured spatial resolution of 4  μm 4  μm . We show simultaneous imaging of neutrophils, monocytes, and fungus (Aspergillus fumigatus) in ex vivo human lung tissue. The instrument has 10 nM and 50 nM sensitivity for fluorescein and Cy5 solutions, respectively. Lung tissue autofluorescence remains visible at up to 200 fps camera acquisition rate. The optical system lends itself to clinical translation due to high-fluorescence sensitivity, simplicity, and the ability to multiplex several pathological molecular imaging targets simultaneously.

Citation

Krstajić, N., Akram, A. R., Choudhary, T. R., McDonald, N., Tanner, M. G., Pedretti, E., …Dhaliwal, K. (2016). Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue. Journal of Biomedical Optics, 21(4), Article 046009. https://doi.org/10.1117/1.jbo.21.4.046009

Journal Article Type Article
Acceptance Date Mar 24, 2016
Online Publication Date Apr 27, 2016
Publication Date Apr 27, 2016
Deposit Date Apr 28, 2016
Publicly Available Date Mar 29, 2024
Journal Journal of Biomedical Optics
Print ISSN 1083-3668
Electronic ISSN 1560-2281
Publisher Society of Photo-optical Instrumentation Engineers
Peer Reviewed Peer Reviewed
Volume 21
Issue 4
Article Number 046009
DOI https://doi.org/10.1117/1.jbo.21.4.046009

Files

Published Journal Article (9.7 Mb)
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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright Statement
© The Authors. Published by SPIE under a Creative Commons
Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including
its DOI. [DOI: 10.1117/1.JBO.21.4.046009]





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