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Related Experiment Video

Updated: May 12, 2026

Second Harmonic Generation Signals in Rabbit Sclera As a Tool for Evaluation of Therapeutic Tissue Cross-linking (TXL) for Myopia
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Characterizing the morphologic changes in collagen crosslinked-treated corneas by Fourier transform-second harmonic

Hsin-Yuan Tan1, Yu-Lin Chang, Wen Lo

  • 1Institute of Biomedical Engineering, College of Medicine and Engineering, National Taiwan University, Taipei, Taiwan.

Journal of Cataract and Refractive Surgery
|April 24, 2013
PubMed
Summary

Forward second harmonic generation (SHG) and 2D-FFT analysis effectively characterize corneal stromal changes after collagen crosslinking (CXL). This label-free method quantifies the shift from linear to wavy collagen fiber patterns post-treatment.

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Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts
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Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts

Published on: October 21, 2022

Area of Science:

  • Ophthalmology
  • Biophysics
  • Biomaterials Science

Background:

  • Corneal collagen crosslinking (CXL) is a therapeutic procedure to strengthen the cornea.
  • Understanding the structural changes in corneal stroma post-CXL is crucial for evaluating treatment efficacy.
  • Label-free imaging techniques offer advantages for characterizing biological tissues without exogenous agents.

Purpose of the Study:

  • To assess the utility of forward second harmonic generation (SHG) imaging combined with 2D-FFT analysis.
  • To achieve label-free characterization and quantification of morphologic alterations in the corneal stroma following CXL.
  • To investigate changes in collagen fiber organization after CXL treatment.

Main Methods:

  • Ex vivo porcine corneas were subjected to forward SHG imaging and 2D-FFT analysis at various depths.
  • Morphologic changes in stromal collagen fibers were evaluated in control and CXL-treated corneas (UVA, riboflavin, or riboflavin-UVA).
  • Histological staining (Hematoxylin-eosin and Sirius red) was used for comparative analysis.

Main Results:

  • Corneal stroma treated with CXL exhibited a transition from linear to wavy collagen fiber patterns.
  • Quantitative 2D-FFT analysis of SHG images revealed increased standard deviations in collagen fiber orientation distribution.
  • These findings indicate altered stromal collagen fiber organization post-CXL.

Conclusions:

  • Forward SHG imaging effectively visualizes morphologic changes in corneal collagen after CXL.
  • Collagen fibers in the corneal stroma change from a linear to a wavy pattern post-CXL.
  • 2D-FFT analysis provides a quantitative method to assess these CXL-induced morphologic changes.