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Cited by 139Advanced Fluorescence Microscopy TechniquesCorneal surgery and disordersSpectroscopy Techniques in Biomedical and Chemical Research

In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

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Latour, Gaël, Gusachenko, Ivan, Kowalczuk, Laura, Lamarre, Isabelle, Schanne-Klein, Marie-Claire·2012·Biomedical Optics Express
DOI 10.1364/boe.3.000001PMC PMC3255328
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Abstract

The transparency and mechanical strength of the cornea are related to the highly organized three-dimensional distribution of collagen fibrils. It is of great interest to develop specific and contrasted in vivo imaging tools to probe these collagenous structures, which is not available yet. Second Harmonic Generation (SHG) microscopy is a unique tool to reveal fibrillar collagen within unstained tissues, but backward SHG images of cornea fail to reveal any spatial features due to the nanometric diameter of stromal collagen fibrils. To overcome this limitation, we performed polarization-resolved SHG imaging, which is highly sensitive to the sub-micrometer distribution of anisotropic structures. Using advanced data processing, we successfully retrieved the orientation of the collagenous fibrils at each depth of human corneas, even in backward SHG homogenous images. Quantitative information was also obtained about the submicrometer heterogeneities of the fibrillar collagen distribution by measuring the SHG anisotropy. All these results were consistent with numerical simulation of the polarization-resolved SHG response of cornea. Finally, we performed in vivo SHG imaging of rat corneas and achieved structural imaging of corneal stroma without any labeling. Epi-detected polarization-resolved SHG imaging should extend to other organs and become a new diagnosis tool for collagen remodeling.

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  • Second harmonic generation imaging can provide qualitative and quantitative information about lamellar organisation within the cornea and has the potential to be used in vivo.

    “Over the last decade there has been an upsurge in the use of non-linear microscopy, in particular second harmonic generation imaging, to provide qualitative (Han et al., 2005, Morishige et al., 2006, Teng et al., 2006, Bueno et al., 2011) and quantitative (Mega et al., 2012, Tuer et al., 2012, Ghazaryan et al., 2013) information about the lamellar organisation within the cornea, a technique that,”

    From Corneal structure and transparency (2015) · cites this paper as Latour et al., 2012

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Authors
Latour, G., Gusachenko, I., Kowalczuk, L., Lamarre, I., & Schanne-Klein, M.-C.
Affiliations
Centre National de la Recherche Scientifique (FR)
ORCID
Venue
Biomedical Optics Express
Publisher
Optica Publishing Group
Published
2011-12-01
Volume
3
Issue
1
Pages
1
ISSN
2156-7085 (ISSN-L)
DOI
10.1364/boe.3.000001
PMC
PMC3255328
OpenAlex
W2105148077
Cited by
139
References
0
Citations by year
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FWCI
5.79
Type
journal article
Language
en
Open access
Licence
Locations
  • Biomedical Optics Express (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, other-oa)
Funders
  • Agence Nationale de la Recherche
  • Fondation de France
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Corneal surgery and disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
Keywords
cornea, collagen fibrils, polarization-resolved second harmonic generation, second harmonic generation microscopy, collagen orientation, corneal stroma, in vivo imaging, label-free imaging, collagen remodeling, rat cornea

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