May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Multiphoton Autofluorescence and Second Harmonic Generation Microscopy for Imaging Corneal Pathologies
Author Affiliations & Notes
  • H.–Y. Tan
    Ophthalmology, Chang–Gung Memorial Hospital, TaoYuan, Taiwan Republic of China
    Institute of Biomedical Engineering, Dermatology,
    National Taiwan University, Taipei, Taiwan Republic of China
  • S.–J. Lin
    Institute of Biomedical Engineering, Dermatology,
    National Taiwan University, Taipei, Taiwan Republic of China
    National Taiwan University Hospital, Taipei, Taiwan Republic of China
  • W. Lo
    Physics, Pathology,
    National Taiwan University, Taipei, Taiwan Republic of China
  • S.–W. Teng
    Physics, Pathology,
    National Taiwan University, Taipei, Taiwan Republic of China
  • R.–J. Wu
    Physics, Pathology,
    National Taiwan University, Taipei, Taiwan Republic of China
  • W.–C. Lin
    Physics, Pathology,
    National Taiwan University Hospital, Taipei, Taiwan Republic of China
  • C.–H. Hsiao
    Ophthalmology, Chang–Gung Memorial Hospital, TaoYuan, Taiwan Republic of China
  • D.–K. Ma
    Ophthalmology, Chang–Gung Memorial Hospital, TaoYuan, Taiwan Republic of China
  • S.–M. Huang
    Ophthalmology, Chang–Gung Memorial Hospital, TaoYuan, Taiwan Republic of China
  • C.–Y. Dong
    Physics, Pathology,
    National Taiwan University, Taipei, Taiwan Republic of China
  • Footnotes
    Commercial Relationships  H. Tan, None; S. Lin, None; W. Lo, None; S. Teng, None; R. Wu, None; W. Lin, None; C. Hsiao, None; D. Ma, None; S. Huang, None; C. Dong, None.
  • Footnotes
    Support  National Research Program for Genomic Medicine, Taiwan (NSC93–3112–B–002–033 and NSC93–3112–B–002–034)
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 2953. doi:
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    • Get Citation

      H.–Y. Tan, S.–J. Lin, W. Lo, S.–W. Teng, R.–J. Wu, W.–C. Lin, C.–H. Hsiao, D.–K. Ma, S.–M. Huang, C.–Y. Dong; Multiphoton Autofluorescence and Second Harmonic Generation Microscopy for Imaging Corneal Pathologies . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2953.

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

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Abstract

Purpose: : To demonstrate the application of multiphoton autofluorescence and second harmonic generation microscopy for imaging corneal pathologies.

Methods: : Human corneal specimens with representative corneal pathologies with different etiologies were obtained during penetrating keratoplasty. A home–built multiphoton microscope was used for imaging the cellular and collagenous structural alterations. Cellular components can be visualized with cytoplasmic autofluorescence, and the collagen fibers can be demonstrated with the non–linear optical effect of second harmonic generation.

Results: : Autofluorescent activated keratocytes and inflammatory cells can be visualized within diseased cornea, which is different from the queiscent autofluorescence–free keratocytes in normal cornea. Destruction and alteration of stroma collagen can also be highlighted with the malalignment or abscence of second harmonic generation signals.

Conclusions: : We demonstrate that multiphoton autofluorescence and second harmonic generation microscopy can provide subcellular resolution for noninvasively imaging corneal pathologies, which may be a powerful tool for corneal physiological and pathological researches, with the potential to be developed into an in vivo imaging modality.

Keywords: microscopy: light/fluorescence/immunohistochemistry • cornea: clinical science • pathology: human 
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