Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation
Abstract Stiffness-related eye diseases such as keratoconus require comprehensive visualization of the complex morphological matrix changes. The aim of this study was to use three-dimensional (3D) light sheet fluorescence microscopy (LSFM) to analyze unlabeled corneal tissue samples, qualitatively v...
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Nature Portfolio
2024-11-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-78516-x |
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| author | Axel Stoecker Diana Pinkert-Leetsch Timea Koch Roland Ackermann Stefan Nolte Christian van Oterendorp Christoph Russmann Jeannine Missbach-Guentner |
| author_facet | Axel Stoecker Diana Pinkert-Leetsch Timea Koch Roland Ackermann Stefan Nolte Christian van Oterendorp Christoph Russmann Jeannine Missbach-Guentner |
| author_sort | Axel Stoecker |
| collection | DOAJ |
| description | Abstract Stiffness-related eye diseases such as keratoconus require comprehensive visualization of the complex morphological matrix changes. The aim of this study was to use three-dimensional (3D) light sheet fluorescence microscopy (LSFM) to analyze unlabeled corneal tissue samples, qualitatively visualizing changes in corneal stiffness. Isolated porcine corneal tissue samples were treated with either NaCl or 0.1% glutaraldehyde (GTA) prior to clearing with benzyl alcohol/benzyl benzoate (BABB) and subsequently scanned with LSFM. After analysis of the LSFM data sets, the samples were embedded in paraffin to validate the results by conventional planar microscopy. In the unlabeled corneal tissue samples the 2D/3D morphology of the entire tissue volume was identified by specific autofluorescence signals. An enhancement of collagen crosslinking was induced by applying GTA to the corneal tissue. Subsequent LSFM scans showed specific morphological changes due to altered autofluorescence signals of the corneal stroma, which were confirmed by conventional histology. Therefore, LSFM analysis of corneal tissue samples allowed label-free 3D autofluorescence assessment of the corneal morphology in its anatomical context. It provides the technical basis for the examination of the pathologically altered cornea and facilitates ophthalmologic examinations of corneal diseases based on the altered tissue stiffness. |
| format | Article |
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| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-fc9781a323e9493f9d5cfa4078012df62024-11-17T12:29:24ZengNature PortfolioScientific Reports2045-23222024-11-0114111010.1038/s41598-024-78516-xCollagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluationAxel Stoecker0Diana Pinkert-Leetsch1Timea Koch2Roland Ackermann3Stefan Nolte4Christian van Oterendorp5Christoph Russmann6Jeannine Missbach-Guentner7Faculty of Engineering and Health, University of Applied Science and ArtsDepartment of Diagnostic and Interventional Radiology, University Medical Center GöttingenInstitute of Applied Physics, Abbe Center of Photonics, Friedrich-Schiller-Universität JenaInstitute of Applied Physics, Abbe Center of Photonics, Friedrich-Schiller-Universität JenaInstitute of Applied Physics, Abbe Center of Photonics, Friedrich-Schiller-Universität JenaDepartment of Ophthalmology, University Medical Center GöttingenFaculty of Engineering and Health, University of Applied Science and ArtsDepartment of Diagnostic and Interventional Radiology, University Medical Center GöttingenAbstract Stiffness-related eye diseases such as keratoconus require comprehensive visualization of the complex morphological matrix changes. The aim of this study was to use three-dimensional (3D) light sheet fluorescence microscopy (LSFM) to analyze unlabeled corneal tissue samples, qualitatively visualizing changes in corneal stiffness. Isolated porcine corneal tissue samples were treated with either NaCl or 0.1% glutaraldehyde (GTA) prior to clearing with benzyl alcohol/benzyl benzoate (BABB) and subsequently scanned with LSFM. After analysis of the LSFM data sets, the samples were embedded in paraffin to validate the results by conventional planar microscopy. In the unlabeled corneal tissue samples the 2D/3D morphology of the entire tissue volume was identified by specific autofluorescence signals. An enhancement of collagen crosslinking was induced by applying GTA to the corneal tissue. Subsequent LSFM scans showed specific morphological changes due to altered autofluorescence signals of the corneal stroma, which were confirmed by conventional histology. Therefore, LSFM analysis of corneal tissue samples allowed label-free 3D autofluorescence assessment of the corneal morphology in its anatomical context. It provides the technical basis for the examination of the pathologically altered cornea and facilitates ophthalmologic examinations of corneal diseases based on the altered tissue stiffness.https://doi.org/10.1038/s41598-024-78516-xExtracellular matrix stiffnessCorneal stromaLight sheet microscopyTomographic microscopyAutofluorescence3D virtual histology |
| spellingShingle | Axel Stoecker Diana Pinkert-Leetsch Timea Koch Roland Ackermann Stefan Nolte Christian van Oterendorp Christoph Russmann Jeannine Missbach-Guentner Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation Scientific Reports Extracellular matrix stiffness Corneal stroma Light sheet microscopy Tomographic microscopy Autofluorescence 3D virtual histology |
| title | Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation |
| title_full | Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation |
| title_fullStr | Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation |
| title_full_unstemmed | Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation |
| title_short | Collagen crosslinking-induced corneal morphological changes: a three-dimensional light sheet Microscopy-based evaluation |
| title_sort | collagen crosslinking induced corneal morphological changes a three dimensional light sheet microscopy based evaluation |
| topic | Extracellular matrix stiffness Corneal stroma Light sheet microscopy Tomographic microscopy Autofluorescence 3D virtual histology |
| url | https://doi.org/10.1038/s41598-024-78516-x |
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