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Optics Letters

Optics Letters


  • Vol. 27, Iss. 23 — Dec. 2, 2002
  • pp: 2082–2084

Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence

Alvin T. Yeh, Nader Nassif, Aikaterini Zoumi, and Bruce J. Tromberg  »View Author Affiliations

Optics Letters, Vol. 27, Issue 23, pp. 2082-2084 (2002)

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A multiphoton microscope employing second-harmonic generation (SHG) and two-photon excited fluorescence (TPF) is used for high-resolution ex vivo imaging of rabbit cornea in a backscattering geometry. Endogenous TPF and SHG signals from corneal cells and extracellular matrix, respectively, are clearly visible without exogenous dyes. Spectral characterization of these upconverted signals provides confirmation of the structural origin of both TPF and SHG, and spectral imaging facilitates the separation of keratocyte and epithelial cells from the collagen-rich corneal stroma. The polarization dependence of collagen SHG is used to highlight fiber orientation, and three-dimensional SHG tomography reveals that approximately 88% of the stromal volume is occupied by collagen lamellae.

© 2002 Optical Society of America

OCIS Codes
(170.5810) Medical optics and biotechnology : Scanning microscopy
(170.6930) Medical optics and biotechnology : Tissue
(300.6410) Spectroscopy : Spectroscopy, multiphoton

Alvin T. Yeh, Nader Nassif, Aikaterini Zoumi, and Bruce J. Tromberg, "Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence," Opt. Lett. 27, 2082-2084 (2002)

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