April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
INVESTIGATING MICROSTRUCTURES OF HUMAN CORNEAL ENDOTHELIAL CELL MICROENVIRONMENT USING HIGH RESOLUTION IMAGING GABOR-DOMAIN OPTICAL COHERENCE MICROSCOPY
Author Affiliations & Notes
  • Patrice Tankam
    Institute of Optics, University of Rochester, Rochester, NY
    Center for Visual Science, University of Rochester, Rochester, NY
  • Jungeun Won
    Department of Biomedical Engineering, University of Rochester, Rochester, NY
  • Anand P Santhanam
    Department of Radiation Oncology, University of California, Los Angeles, CA
  • Zhiguo He
    Corneal Graft Biology, Engineering and Imaging Laboratory, EA 2125, SFR143, Faculty of Medicine, Jean Monnet University, Saint Etienne, France
  • Pataia Giacomo
    Corneal Graft Biology, Engineering and Imaging Laboratory, EA 2125, SFR143, Faculty of Medicine, Jean Monnet University, Saint Etienne, France
    Institut d'Optique (site of Saint-Etienne) and Université de Lyon, CNRS, Laboratoire Hubert Curien, Jean Monnet University, Saint Etienne, France
  • Gain Philippe
    Corneal Graft Biology, Engineering and Imaging Laboratory, EA 2125, SFR143, Faculty of Medicine, Jean Monnet University, Saint Etienne, France
  • Gilles Thuret
    Corneal Graft Biology, Engineering and Imaging Laboratory, EA 2125, SFR143, Faculty of Medicine, Jean Monnet University, Saint Etienne, France
  • Thierry Lepine
    Institut d'Optique (site of Saint-Etienne) and Université de Lyon, CNRS, Laboratoire Hubert Curien, Jean Monnet University, Saint Etienne, France
  • Holly Butler Hindman
    Center for Visual Science, University of Rochester, Rochester, NY
    Department of Ophthalmology, University of Rochester Medical Center, School of Medicine and Dentistry, Rochester, NY
  • Jannick Rolland
    Institute of Optics, University of Rochester, Rochester, NY
    Center for Visual Science, University of Rochester, Rochester, NY
  • Footnotes
    Commercial Relationships Patrice Tankam, None; Jungeun Won, None; Anand Santhanam, None; Zhiguo He, None; Pataia Giacomo, None; Gain Philippe, None; Gilles Thuret, None; Thierry Lepine, None; Holly Hindman, None; Jannick Rolland, LighTopTech Corp (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 2071. doi:
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      Patrice Tankam, Jungeun Won, Anand P Santhanam, Zhiguo He, Pataia Giacomo, Gain Philippe, Gilles Thuret, Thierry Lepine, Holly Butler Hindman, Jannick Rolland, ; INVESTIGATING MICROSTRUCTURES OF HUMAN CORNEAL ENDOTHELIAL CELL MICROENVIRONMENT USING HIGH RESOLUTION IMAGING GABOR-DOMAIN OPTICAL COHERENCE MICROSCOPY. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2071.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

We propose here to use a high volumetric resolution imaging modality, Gabor-Domain Optical Coherence Microscopy (GD-OCM), to observe corneal endothelial cells and corneal microstructures in order to better understand corneal layers functionality

 
Methods
 

GD-OCM combines high sectioning capability of optical coherence tomography (OCT) and high lateral resolution of confocal microscopy to achieve high-contrast imaging with volumetric cellular resolution of 2 μm across a thickness of up to 2 mm in tissue. The current GD-OCM system fits on a movable cart. The handheld scanning probe is attached to an articulated arm that can be easily adjusted. The light source is a superluminescent diode laser centered at 840 nm with 100 nm FWHM (BroadLighter D-840-HP-I, Superlum®, Ireland). The microscope objective probe incorporates a liquid lens, which allows dynamic-focusing in order to image different depths of the sample. A custom dispersion compensator and a custom spectrometer with a high-speed CMOS line camera (spl4096-70km, Basler Inc.) are used to acquire the spectral information. We recently implemented and will also present a parallelized Multi-Graphic Processing Units (GPU) architecture of the system to allow real-time visualization of the sample. The overall time to 3D visualization, including acquisition, processing and rendering of a 1000 × 1000 × 400 voxels, is less than 2 minutes compared to 5 hours on a conventional CPU. The lateral resolution of 2 µm (significantly far from conventional OCT) coupled to the high axial resolution of 2µm allowed analyzing the en face and cross sectional imaging of corneal layers. This imaging modality, with 1 x 1 mm2 field of view and 1 mm imaging depth in corneal tissue, was devoted to investigate the microenvironment of excised human corneas

 
Results
 

The imaging system revealed high contrast images of stroma fibers and the distribution of endothelial cell nuclei. Cross-section x-z images also allow the visualization of stroma keratocytes

 
Conclusions
 

This new imaging modality revealed key features of the corneal layers. Future work will consist of studying these features in order to correlate their morphology to their functionality. This work will provide insights into the mechanism of corneal diseases and particularly endothelial cells dysfunction

  
Keywords: 481 cornea: endothelium • 552 imaging methods (CT, FA, ICG, MRI, OCT, RTA, SLO, ultrasound) • 494 degenerations/dystrophies  
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