Multimodal, multiphoton microscopy and image correlation analysis for characterizing corneal thermal damage.
We used the combination of multiphoton autofluorescence (MAF), forward second-harmonic generation (FWSHG), and backward second-harmonic generation (BWSHG) imaging for the qualitative and quantitative characterization of thermal damage of ex vivo bovine cornea. We attempt to characterize the structural alterations by qualitative MAF, FWSHG, and BWSHG imaging in the temperature range of 37 to 90 degrees C. In addition to measuring the absolute changes in the three types of signals at the stromal surface, we also performed image correlation analysis between FWSHG and BWSHG and demonstrate that with increasing thermal damage, image correlation between FWSHG and BWSHG significantly increases. Our results show that while MAF and BWSHG intensities may be used as preliminary indicators of the extent of corneal thermal damage, the most sensitive measures are provided by the decay in FWSHG intensity and the convergence of FWSHG and BWSHG images.
Lo
Wen
W
National Taiwan University, Department of Physics and Center for Quantum Science and Engineering, Tainan, Taiwan.
Chang
Yu-Lin
YL
Liu
Jia-Shiu
JS
Hseuh
Chiu-Mei
CM
Hovhannisyan
Vladimir
V
Chen
Shean-Jen
SJ
Tan
Hsin-Yuan
HY
Dong
Chen-Yuan
CY
D000818:Animals; D002417:Cattle; D003315:Cornea / Q000473:pathology*; D065306:Corneal Injuries; D005126:Eye Burns / Q000473:pathology*; D007089:Image Enhancement / Q000379:methods*; D066298:In Vitro Techniques; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013382:Subtraction Technique