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

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

doi:10.1117/1.JBO.21.4.046009

27 April 2016 Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue

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

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Journal of Biomedical Optics, Vol. 21, Issue 4, 046009 (April 2016). https://doi.org/10.1117/1.JBO.21.4.046009

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). This effectively increases the measured spatial resolution of 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.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Nikola Krstajić, Ahsan R. Akram M.D., Tushar R. Choudhary, Neil McDonald, Michael G. Tanner, Ettore Pedretti, Paul A. Dalgarno, Emma Scholefield, John M. Girkin, Anne Moore, Mark Bradley, and Kevin Dhaliwal M.D. "Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue," Journal of Biomedical Optics 21(4), 046009 (27 April 2016). https://doi.org/10.1117/1.JBO.21.4.046009

Published: 27 April 2016

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