High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria

Ettore Pedretti, Michael George Tanner, Tushar R. Choudhary, Nikola Krstajić, Alicia Megia-Fernandez, Robert K. Henderson, Mark Bradley, Robert R. Thomson, John M. Girkin, Kevin Dhaliwal, Paul A. Dalgarno

Research output: Contribution to journalArticle

Abstract

We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence imaging by temporally shifting the dual excitation lasers, with respect to each other, to separate the two spectrally distinct fluorescent decays in time. Combining the temporal encoding, to provide spectral separation, with lifetime measurements we show a one FPS, multi-channel endomicroscopy platform for clinical applications and diagnosis. We demonstrate the potential of the system by imaging SmartProbe labeled bacteria in ex vivo samples of human lung using lifetimeto differentiate bacterial fluorescence from the strong background lung autofluorescence which was used to provide structural information.
LanguageEnglish
Pages181-195
Number of pages15
JournalBiomedical Optics Express
Volume10
Issue number1
Early online date12 Dec 2018
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

bacteria
high speed
color
life (durability)
fluorescence
lungs
photons
avalanche diodes
scanning
distributing
piles
excitation
lasers
coding
platforms
fibers
detectors
decay
pulses

Keywords

  • Laser scanning
  • Laser sources
  • Photon counting
  • Spatial light modulators
  • Spectral imaging
  • Ti:sapphire lasers

Cite this

Pedretti, Ettore ; Tanner, Michael George ; Choudhary, Tushar R. ; Krstajić, Nikola ; Megia-Fernandez, Alicia ; Henderson, Robert K. ; Bradley, Mark ; Thomson, Robert R. ; Girkin, John M. ; Dhaliwal, Kevin ; Dalgarno, Paul A. / High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria. In: Biomedical Optics Express. 2019 ; Vol. 10, No. 1. pp. 181-195.
@article{58432b379e13469692ea6ee47815586c,
title = "High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria",
abstract = "We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence imaging by temporally shifting the dual excitation lasers, with respect to each other, to separate the two spectrally distinct fluorescent decays in time. Combining the temporal encoding, to provide spectral separation, with lifetime measurements we show a one FPS, multi-channel endomicroscopy platform for clinical applications and diagnosis. We demonstrate the potential of the system by imaging SmartProbe labeled bacteria in ex vivo samples of human lung using lifetimeto differentiate bacterial fluorescence from the strong background lung autofluorescence which was used to provide structural information.",
keywords = "Laser scanning, Laser sources, Photon counting, Spatial light modulators, Spectral imaging, Ti:sapphire lasers",
author = "Ettore Pedretti and Tanner, {Michael George} and Choudhary, {Tushar R.} and Nikola Krstajić and Alicia Megia-Fernandez and Henderson, {Robert K.} and Mark Bradley and Thomson, {Robert R.} and Girkin, {John M.} and Kevin Dhaliwal and Dalgarno, {Paul A.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1364/BOE.10.000181",
language = "English",
volume = "10",
pages = "181--195",
journal = "Biomedical Optics Express",
issn = "2156-7085",
publisher = "The Optical Society",
number = "1",

}

High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria. / Pedretti, Ettore; Tanner, Michael George; Choudhary, Tushar R.; Krstajić, Nikola; Megia-Fernandez, Alicia; Henderson, Robert K.; Bradley, Mark; Thomson, Robert R.; Girkin, John M.; Dhaliwal, Kevin; Dalgarno, Paul A.

In: Biomedical Optics Express, Vol. 10, No. 1, 01.01.2019, p. 181-195.

Research output: Contribution to journalArticle

TY - JOUR

T1 - High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria

AU - Pedretti, Ettore

AU - Tanner, Michael George

AU - Choudhary, Tushar R.

AU - Krstajić, Nikola

AU - Megia-Fernandez, Alicia

AU - Henderson, Robert K.

AU - Bradley, Mark

AU - Thomson, Robert R.

AU - Girkin, John M.

AU - Dhaliwal, Kevin

AU - Dalgarno, Paul A.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence imaging by temporally shifting the dual excitation lasers, with respect to each other, to separate the two spectrally distinct fluorescent decays in time. Combining the temporal encoding, to provide spectral separation, with lifetime measurements we show a one FPS, multi-channel endomicroscopy platform for clinical applications and diagnosis. We demonstrate the potential of the system by imaging SmartProbe labeled bacteria in ex vivo samples of human lung using lifetimeto differentiate bacterial fluorescence from the strong background lung autofluorescence which was used to provide structural information.

AB - We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence imaging by temporally shifting the dual excitation lasers, with respect to each other, to separate the two spectrally distinct fluorescent decays in time. Combining the temporal encoding, to provide spectral separation, with lifetime measurements we show a one FPS, multi-channel endomicroscopy platform for clinical applications and diagnosis. We demonstrate the potential of the system by imaging SmartProbe labeled bacteria in ex vivo samples of human lung using lifetimeto differentiate bacterial fluorescence from the strong background lung autofluorescence which was used to provide structural information.

KW - Laser scanning

KW - Laser sources

KW - Photon counting

KW - Spatial light modulators

KW - Spectral imaging

KW - Ti:sapphire lasers

U2 - 10.1364/BOE.10.000181

DO - 10.1364/BOE.10.000181

M3 - Article

VL - 10

SP - 181

EP - 195

JO - Biomedical Optics Express

T2 - Biomedical Optics Express

JF - Biomedical Optics Express

SN - 2156-7085

IS - 1

ER -