Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy

In this study, we use second harmonic generation (SHG) microscopy to visualize the collagen distribution over the full depth of 10 intact and unstained human corneas (500-600 µm thick). We take advantage of the small coherence length in epi-detection to axially resolve the lamellae while maintaining the corneal physiological curvature. Moreover, as raw epi-detected SHG images are spatially homogenous because of the sub-wavelength size of stromal collagen fibrils, we use a polarimetric approach to measure the collagen orientation in every voxel. After a careful validation of this approach, we show that the collagen lamellae (i) are mostly oriented along the inferior-superior axis in the anterior stroma and along the nasal-temporal axis in the posterior stroma, with a gradual shift in between and (ii) exhibit more disorder in the anterior stroma. These results represent the first quantitative characterization of the lamellar structure of the human cornea continuously along its entire thickness with micrometric resolution. It also shows the unique potential of P-SHG microscopy for imaging of collagen distribution in thick dense tissues.PMID:37528091 | DOI:10.1038/s41377-023-01224-0

Source: Appl Human Sci - August 1, 2023 Category: Physiology Authors: Clothilde Raoux Anatole Chessel Pierre Mahou Ga ël Latour Marie-Claire Schanne-Klein Source Type: research