Quantitative analysis of thermally-induced alterations of corneal stroma by second-harmonic generation imaging

Thermal modifications induced in the corneal stroma were investigated by means of second harmonic generation (SHG) imaging. Whole fresh cornea samples were heated in a water bath at temperatures in the 35-80 degrees C range for a 4-min time. SHG images of the structural modifications induced at each temperature were acquired from different areas of cross-sectioned corneal stroma by using an 880 nm linearly- and circularly-polarized excitation light emitted by a mode-locked Ti:Sapphire laser. The SHG images were then analyzed by means of both an empirical approach and a 2D-theoretical model. The proposed analyses provide a detailed description of the changes occurring in the structural architecture of the cornea during the thermal treatment. Our results allow us to depict a temperature-dependent biochemical model for the progressive destructuration occurring to collagen fibrils and nonfibrillar components of the stroma.