In vitro characterization of corneal wound healing using multiphoton autofluorescence and second harmonic generation (SHG) microscopy

Yen Sun; Wen Lo; Wei-Liang Chen; Shu-Wen Teng; Hsin-Yuan Tan; Chen-Yuan Dong

doi:10.1117/12.647413

7 March 2006 In vitro characterization of corneal wound healing using multiphoton autofluorescence and second harmonic generation (SHG) microscopy

Yen Sun, Wen Lo, Wei-Liang Chen, Shu-Wen Teng, Hsin-Yuan Tan, Chen-Yuan Dong

Author Affiliations +

Proceedings Volume 6138, Ophthalmic Technologies XVI; 61381U (2006) https://doi.org/10.1117/12.647413
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

The purpose of this investigation is to characterize corneal wound healing under in vitro conditions. Multiphoton autofluorescence and second harmonic generation (SHG) microscopy will be used to visualize cells and collagen fibers associated with corneal wound healing. Using the near-infrared excitation source from a titanium-sapphire laser pumped by a diode-pumped, solid state (DPSS) laser system, we can induce and simultaneously acquire multiphoton autofluorescence and SHG signals from the cornea specimens. A home-modified commercial microscope system with specified optical components is used for optimal signal detection. To acquire both high resolution and tissue-level information of the specimen, a sample positioning stage is used in conjunction with the beam scanning system. Finally, the organ level image can be assembled from individual area scans. The in vitro samples we used are cornea buttons acquired from porcine eyes. Localized wounds will be induced by #11 blade and imaged using multiphoton microscopy. Based on these results, we envision the in vitro imaging chamber to be able to follow the wound healing process without damaging histological procedures. We envision this approach will enable us to further understand wound healing process associated with corneal scar and can lead to in vivo methodology for diagnosing cornea damage.

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Yen Sun, Wen Lo, Wei-Liang Chen, Shu-Wen Teng, Hsin-Yuan Tan, and Chen-Yuan Dong "In vitro characterization of corneal wound healing using multiphoton autofluorescence and second harmonic generation (SHG) microscopy", Proc. SPIE 6138, Ophthalmic Technologies XVI, 61381U (7 March 2006); https://doi.org/10.1117/12.647413

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KEYWORDS

Second-harmonic generation

Wound healing

Cornea

Multiphoton microscopy

Microscopy

In vitro testing

Multiphoton fluorescence microscopy

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