Two-color widefield fluorescence microendoscopy enables multiplexed molecular imaging in the alveolar space of human lung tissue.
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.
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United KingdombUniversity of Edinburgh, School of E.
Nikola
N
Krstajic
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Ahsan R
AR
Akram
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United KingdomcHeriot-Watt University, Institute of.
Tushar R
TR
Choudhary
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Neil
N
McDonald
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United KingdomdHeriot-Watt University, Institute of.
Michael G
MG
Tanner
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United KingdomcHeriot-Watt University, Institute of.
Ettore
E
Pedretti
Heriot-Watt University, Institute of Biological Chemistry, Biophysics and Bioengineering, Edinburgh EH14 4AS, United Kingdom.
Paul A
PA
Dalgarno
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Emma
E
Scholefield
Durham University, Biophysical Sciences Institute, Department of Physics, South Road, Durham DH1 3LE, United Kingdom.
John M
JM
Girkin
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Anne
A
Moore
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Mark
M
Bradley
University of Edinburgh, Queen's Medical Research Institute, MRC Centre for Inflammation Research, EPSRC IRC "Hub" in Optical Molecular Sensing and Imaging, 47 Little France Crescent, Edinburgh EH16 4TJ, United Kingdom.
Kevin
K
Dhaliwal
D001232:Aspergillus fumigatus / Q000737:chemistry; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D004867:Equipment Design; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008168:Lung / Q000166:cytology* / Q000382:microbiology*; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D009000:Monocytes / Q000166:cytology; D009504:Neutrophils / Q000166:cytology; D011550:Pseudomonas aeruginosa / Q000737:chemistry