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

Nikola Krstajic (Lead / Corresponding author), Ahsan R Akram, Tushar R Choudhary, Neil McDonald, Michael G Tanner, Ettore Pedretti, Paul A Dalgarno, Emma Scholefield, John M Girkin, Anne Moore, Mark Bradley, Kevin Dhaliwal (Lead / Corresponding author)

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    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.

    Original languageEnglish
    Article number46009
    Number of pages13
    JournalJournal of Biomedical Optics
    Volume21
    Issue number4
    DOIs
    Publication statusPublished - 30 Apr 2016

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    Keywords

    • Aspergillus fumigatus
    • Bronchoalveolar Lavage Fluid
    • Equipment Design
    • Humans
    • Image Processing, Computer-Assisted
    • Lung
    • Microscopy, Fluorescence
    • Molecular Imaging
    • Monocytes
    • Neutrophils
    • Pseudomonas aeruginosa
    • Journal Article
    • Research Support, Non-U.S. Gov't

    Cite this

    Krstajic, N., Akram, A. R., Choudhary, T. R., McDonald, N., Tanner, M. G., Pedretti, E., Dalgarno, P. A., Scholefield, E., Girkin, J. M., Moore, A., Bradley, M., & 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), [46009]. https://doi.org/10.1117/1.JBO.21.4.046009