May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Second Harmonic Imaging Microscopy of Normal Human and Keratoconus Cornea
N. Morishige; A. J. Wahlert; M. C. Kenney; D. J. Brown; K. Kawamoto; T.-I. Chikama; T. Nishida; J. V. Jester
Author Affiliations & Notes
  • N. Morishige
    Dept. of Ophthalmology, Yamaguchi Univ Grad Sch of Med, Ube, Japan
    The Eye Institute, University of California, Irvine, Orange, California
  • A. J. Wahlert
    The Eye Institute, University of California, Irvine, Orange, California
  • M. C. Kenney
    The Eye Institute, University of California, Irvine, Orange, California
  • D. J. Brown
    The Eye Institute, University of California, Irvine, Orange, California
  • K. Kawamoto
    Dept. of Ophthalmology, Yamaguchi Univ Grad Sch of Med, Ube, Japan
  • T.-I. Chikama
    Dept. of Ophthalmology, Yamaguchi Univ Grad Sch of Med, Ube, Japan
  • T. Nishida
    Dept. of Ophthalmology, Yamaguchi Univ Grad Sch of Med, Ube, Japan
  • J. V. Jester
    The Eye Institute, University of California, Irvine, Orange, California
  • Footnotes
    Commercial Relationships N. Morishige, None; A.J. Wahlert, None; M.C. Kenney, None; D.J. Brown, None; K. Kawamoto, None; T. Chikama, None; T. Nishida, None; J.V. Jester, None.
  • Footnotes
    Support NIH grants EY07348 (Jester), EY016663, Support Grant from Research to Prevent Blindness, Inc, The Skirball Program in Molecular Ophthalmology, and the Japan Eye Bank Association
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 3517. doi:
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      N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T.-I. Chikama, T. Nishida, J. V. Jester; Second Harmonic Imaging Microscopy of Normal Human and Keratoconus Cornea. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3517.

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Abstract

Purpose:: The purpose of this study was to evaluate the ability of second harmonic imaging to identify differences in corneal stromal collagen organization between normal human and keratoconus corneas.

Methods:: Six normal corneas from eye bank donors and 13 corneas obtained from keratoconus patients obtained after penetrating keratoplasty were examined. A femtosecond titanium-sapphire laser with 800 nm output was used to generate second harmonic signals collected at 400 nm from central and paracentral corneal tissue blocks. 3-Dimensional data sets were collected and reconstructed to evaluate the location and orientation of stromal collagen lamellae.

Results:: Imaging of second harmonic signals combined with 3D reconstruction of the normal cornea identified a high degree of lamellar interweaving particularly in the anterior cornea. Of note was the detection of lamellae that inserted into Bowman’s Layer and were oriented transverse to the corneal surface, penetrating posteriorly approximately 120 µm. In keratoconus corneas, imaging second harmonic signals identified less lamellar interweaving and a marked reduction or loss of lamellae inserting into Bowman’s layer in 12 of 13 cases, particularly in regions associated with cone development without breaks in Bowman’s layer or scarring.

Conclusions:: Compared to normal adult corneas, marked abnormalities were detected in the organization of the anterior corneal collagen lamellae of keratoconus corneas by second harmonic imaging. These structural abnormalities are consistent with the known changes in collagen organization and biomechanical strength of keratoconus.

Keywords: cornea: stroma and keratocytes • cornea: clinical science • imaging methods (CT, FA, ICG, MRI, OCT, RTA, SLO, ultrasound) 
© 2007, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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