Multiphoton fluorescence and second-harmonic-generation microscopy for imaging structural alterations in corneal scar tissue in penetrating full-thickness wound.

The transparency of the cornea relies on the special spatial arrangement of stromal collagen. Any pathological abnormality that leads to the wound healing responses and consequential disruption of collagen alignment may hinder corneal transparency. Understanding wound healing may be crucial for the successful applications of clinical procedures such as refractive surgery. Therefore, the development of a noninvasive in vivo imaging technique is valuable for investigating the physiological response associated with corneal wound healing. In this article, we demonstrate multiphoton ex vivo imaging of full-thickness corneal scar tissue 10 years following wounding. We intend to demonstrate the ability of multiphoton imaging to reveal corneal structural alterations from the wound healing process and the potential of this imaging modality in clinical diagnosis and monitoring of corneal pathological abnormalities.