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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 18, Iss. 5 — Mar. 1, 2010
  • pp: 5028–5040

Harmonic microscopy of isotropic and anisotropic microstructure of the human cornea

Nicolas Olivier, Florent Aptel, Karsten Plamann, Marie-Claire Schanne-Klein, and Emmanuel Beaurepaire  »View Author Affiliations

Optics Express, Vol. 18, Issue 5, pp. 5028-5040 (2010)

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In this study we present combined third-harmonic generation (THG) and second-harmonic generation (SHG) microscopy images of intact human corneas, and we analyze experimentally and theoretically the origin of the THG signal. Multiharmonic microscopy provides detailed images of the cornea microstructure over its entire thickness. A component of the THG signal originates from cellular structures and another one originates from anisotropy changes between successive collagen lamellae in the stroma. This anisotropy-related signal can be specifically detected using circular incident polarization, and provide contrasted images of the stacking and tissue-scale heterogeneity of stromal lamellae. Forward-radiated THG and SHG signals are generally anticorrelated, indicating that maximum THG is obtained from lamellar interfaces whereas maximum SHG is obtained from within lamellae. Polarization-resolved THG imaging reflects the alternate anisotropy directions of the lamellae. We present a model for THG imaging of layered anisotropic samples and numerical calculations that account for our observations.

© 2010 Optical Society of America

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.4470) Medical optics and biotechnology : Ophthalmology
(190.4160) Nonlinear optics : Multiharmonic generation
(190.4710) Nonlinear optics : Optical nonlinearities in organic materials
(180.4315) Microscopy : Nonlinear microscopy
(170.6935) Medical optics and biotechnology : Tissue characterization

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: January 20, 2010
Manuscript Accepted: February 11, 2010
Published: February 25, 2010

Virtual Issues
Vol. 5, Iss. 6 Virtual Journal for Biomedical Optics

Nicolas Olivier, Florent Aptel, Karsten Plamann, Marie-Claire Schanne-Klein, and Emmanuel Beaurepaire, "Harmonic microscopy of isotropic and anisotropic microstructure of the human cornea," Opt. Express 18, 5028-5040 (2010)

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