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

Nikola Krstajic, Ahsan R. Akram, Tushar Choudhary, Neil Mcdonald, Michael George Tanner, Ettore Pedretti, Paul Dalgarno, Emma Scholefield, John M. Girkin, Anne Moore, Mark Bradley, Kevin Dhaliwal

Research output: Contribution to journalArticle

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  μm4  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 number046009
JournalJournal of Biomedical Optics
Volume21
Issue number4
DOIs
Publication statusPublished - 27 Apr 2016

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Molecular Imaging
Color
Fluorescence
Lung
Bacteria
Light
Blinking
Optical Devices
Aspergillus fumigatus
Fluorescein
Pseudomonas aeruginosa
Monocytes
Neutrophils
Fungi

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Krstajic, Nikola ; Akram, Ahsan R. ; Choudhary, Tushar ; Mcdonald, Neil ; Tanner, Michael George ; Pedretti, Ettore ; Dalgarno, Paul ; 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. In: Journal of Biomedical Optics. 2016 ; Vol. 21, No. 4.
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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  μm4  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.",
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Krstajic, N, Akram, AR, Choudhary, T, Mcdonald, N, Tanner, MG, Pedretti, E, Dalgarno, P, Scholefield, E, Girkin, JM, 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, vol. 21, no. 4, 046009. https://doi.org/10.1117/1.JBO.21.4.046009

Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue. / Krstajic, Nikola; Akram, Ahsan R.; Choudhary, Tushar; Mcdonald, Neil; Tanner, Michael George; Pedretti, Ettore; Dalgarno, Paul; Scholefield, Emma; Girkin, John M.; Moore, Anne; Bradley, Mark; Dhaliwal, Kevin.

In: Journal of Biomedical Optics, Vol. 21, No. 4, 046009, 27.04.2016.

Research output: Contribution to journalArticle

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AU - Krstajic, Nikola

AU - Akram, Ahsan R.

AU - Choudhary, Tushar

AU - Mcdonald, Neil

AU - Tanner, Michael George

AU - Pedretti, Ettore

AU - Dalgarno, Paul

AU - Scholefield, Emma

AU - Girkin, John M.

AU - Moore, Anne

AU - Bradley, Mark

AU - Dhaliwal, Kevin

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AB - 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  μm4  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.

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