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

Clothilde Raoux; Anatole Chessel; Pierre Mahou; Gaël Latour; Marie-Claire Schanne-Klein

doi:10.1038/s41377-023-01224-0

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

Original Articles

- Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy
- Light: Science & Applications Vol. 12, Issue 9, Pages: 1782-1794(2023)
- Affiliations：
  
  1.Laboratory for Optics and Biosciences, Ecole Polytechnique, CNRS, INSERM, Institut Polytechnique de Paris, 91128 Palaiseau, France
  2.Université Paris-Saclay, 91190 Gif-sur-Yvette, France
- Author bio：
  
  Marie-Claire Schanne-Klein (marie-claire.schanne-klein@polytechnique.edu)
- Funds：
- DOI：10.1038/s41377-023-01224-0
  CLC：
- Published：30 September 2023，
  
  Published Online：02 August 2023，
  
  Received：07 April 2023，
  
  Revised：22 June 2023，
  
  Accepted：05 July 2023
- Accepted：
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Raoux, C. et al. Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy. Light: Science & Applications, 12, 1782-1794 (2023).
DOI：

Raoux, C. et al. Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy. Light: Science & Applications, 12, 1782-1794 (2023). DOI： 10.1038/s41377-023-01224-0.

摘要

Abstract

A key property of the human cornea is to maintain its curvature and consequently its refraction capability despite daily changes in intraocular pressure. This is closely related to the multiscale structure of the corneal stroma

which consists of 1–3 µm-thick stacked lamellae made of thin collagen fibrils. Nevertheless

the distribution

size

and orientation of these lamellae along the depth of the cornea are poorly characterized up to now. 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.

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references

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