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Journal of Biomedical Optics • Open Access

Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue
Author(s): Nikola Krstajić; 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

Paper 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.

Paper Details

Date Published: 27 April 2016
PDF: 13 pages
J. Biomed. Opt. 21(4) 046009 doi: 10.1117/1.JBO.21.4.046009
Published in: Journal of Biomedical Optics Volume 21, Issue 4
Show Author Affiliations
Nikola Krstajić, The Univ. of Edinburgh (United Kingdom)
Ahsan R. Akram, The Univ. of Edinburgh (United Kingdom)
Tushar R. Choudhary, The Univ. of Edinburgh (United Kingdom)
Heriot-Watt Univ. (United Kingdom)
Neil McDonald, The Univ. of Edinburgh (United Kingdom)
Michael G. Tanner, The Univ. of Edinburgh (United Kingdom)
Heriot-Watt Univ. (United Kingdom)
Ettore Pedretti, The Univ. of Edinburgh (United Kingdom)
Heriot-Watt Univ. (United Kingdom)
Paul A. Dalgarno, Heriot-Watt Univ. (United Kingdom)
Emma Scholefield, The Univ. of Edinburgh (United Kingdom)
John M. Girkin, Durham Univ. (United Kingdom)
Anne Moore, The Univ. of Edinburgh (United Kingdom)
Mark Bradley, The Univ. of Edinburgh (United Kingdom)
Kevin Dhaliwal, The Univ. of Edinburgh (United Kingdom)


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