Evaluating Corneal Collagen Organization Using High-Resolution Nonlinear Optical Macroscopy
Open accessAbstract
PURPOSE: Recent developments in nonlinear optical (NLO) imaging using femtosecond lasers provides a noninvasive method for detecting collagen fibers by imaging second harmonic-generated (SHG) signals. However, this technique is limited by the small field of view necessary to generate SHG signals. The purpose of this report is to review our efforts to greatly extend the field of view to assess the entire collagen structure using high-resolution macroscopic (HRMac) imaging. METHODS: Intact human eyes were fixed under pressure, and the whole cornea (13-mm diameter) was excised and embedded in low-melting point agar for vibratome sectioning (200-300 microm). Sections were then optically scanned using a Zeiss LSM 510 Meta and Chameleon femtosecond laser (Carl Zeiss Microimaging Inc., Thornwood, NY) to generate SHG images. For each vibratome section, an overlapping series of three-dimensional data sets (466 x 466 x 150 microm) were taken, covering the entire tissue (15 mm x 6 mm area) using a motorized, mechanical stage. The three-dimensional data sets were then concatenated to generate an NLO-based tomograph. RESULTS: The HRMac of the cornea yielded large macroscopic (80 megapixels per plane), three-dimensional tomographs with high resolution (0.81 microm lateral, 2.0 microm axial) in which individual collagen fibers (stromal lamellae) could be traced, segmented, and extracted. Three-dimensional reconstructions suggested that the anterior cornea comprises highly intertwined lamellae that insert into the anterior limiting lamina (Bowman's layer). CONCLUSIONS: We conclude that HRMac using NLO-based tomography provides a powerful new tool to assess collagen structural organization within the cornea.
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In human corneas, the lamellar organisation through the full thickness from limbus-to-limbus has been reconstructed, demonstrating interweaving of anterior lamellae and their bow spring-like insertions in and out of Bowman's layer.
“Also in human corneas, Jester et al. (2010) were able to reconstruct the lamellar organisation through the full thickness of the cornea from limbus-to-limbus. The reconstructions demonstrated the interweaving of the anterior lamellae and their bow spring-like insertions in and out of Bowman's layer (Fig. 1d).”
From Corneal structure and transparency (2015) · cites this paper as Jester et al., 2010
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Cites
X-Ray Scattering Used to Map the Preferred Collagen Orientation in the Human Cornea and Limbus
Aghamohammadzadeh, H., Newton, R. H., & Meek, K. M. (2004) · Structure · doi:10.1016/j.str.2004.01.002 · Cited by 233 · see citing works
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Corneal structure and transparency
Meek, K. M., & Knupp, C. (2015) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2015.07.001 · Cited by 853 · see citing works
- Authors
- Jester, J. V., Winkler, M., Jester, B. E., Nien, C., Chai, D., & Brown, D. J.
- Affiliations
- University of California, Irvine Medical Center (US)
- Corresponding author
- Jester, James V.
- Jester, Bryan E.
- Venue
- Eye & Contact Lens: Science & Clinical Practice
- Published
- 2010-09
- Volume
- 36
- Issue
- 5
- Pages
- 260-264
- ISSN
- 0733-8902 (ISSN-L)
- 1542-2321
- PMC
- PMC2945240
- OpenAlex
- W2074781029
- Cited by
- 64
- References
- 1
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 5 · 2023: 6 · 2022: 2 · 2021: 2 · 2020: 4 · 2019: 1 · 2018: 7 · 2017: 5 · 2016: 3 · 2015: 6 · 2014: 1 · 2013: 6 · 2012: 3
- FWCI
- 3.99
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Eye & Contact Lens Science & Clinical Practice (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Eye Institute (R01EY007348, EY018665, EY016663, EY07348, R24EY016663, R01EY018665)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Corneal surgery and disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Engineering › Biomedical Engineering › Optical Coherence Tomography Applications
- Keywords
- second harmonic generation, corneal stroma, 3D tomography, Bowman's layer
- MeSH terms
- Collagen, Cornea, Humans, Image Processing, Computer-Assisted, Lasers, Imaging, Three-Dimensional, Tomography, Optical
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