Identifying crossing collagen fibers in human corneal tissues using pSHG images

Polarization sensitive second harmonic generation (pSHG) microscopy has been used previously to characterize the structure of collagen fibers in conical samples. Due to the typical organization of the conical stroma, the information that pSHG provides may be misleading in points where two different collagen fiber bundles orient along different direction crossings. Here a simulation that illustrates the problem is presented. along with a novel method that is capable of identifying these crossing points. These results can be used to improve the evaluation of corneal collagen structure. and it has been applied to analyze pSHG data acquired from healthy and keratocomc human corneal samples. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement