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Evaluating corneal collagen organization using high-resolution nonlinear optical macroscopy.

James V Jester¹, Moritz Winkler, Bryan E Jester

¹Department of Ophthalmology, Gavin Herbert Eye Institute, University of California Irvine Medical Center, Orange, CA, USA. Jjester@uci.edu

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|August 21, 2010

Summary

High-resolution macroscopic imaging with nonlinear optical tomography greatly expands the field of view for detecting collagen fibers in the cornea. This advanced technique allows detailed assessment of the entire collagen structure, offering new insights into corneal organization.

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Area of Science:

Biomedical Optics
Ophthalmology
Materials Science

Background:

Nonlinear optical (NLO) imaging with femtosecond lasers detects collagen fibers via second harmonic-generated (SHG) signals.
Current NLO imaging techniques are limited by a small field of view, hindering comprehensive collagen structure assessment.

Purpose of the Study:

To review efforts in extending the field of view for NLO imaging.
To assess entire collagen structures using high-resolution macroscopic (HRMac) imaging.

Main Methods:

Excised human corneas were sectioned using vibratome.
Optical scanning generated overlapping 3D datasets using a femtosecond laser for SHG imaging.
Datasets were concatenated to create NLO-based tomographs.

Main Results:

HRMac yielded large, high-resolution 3D tomographs of the cornea.
Individual collagen fibers (stromal lamellae) were traceable and extractable.
Reconstructions revealed intertwined lamellae inserting into Bowman's layer.

Conclusions:

HRMac using NLO-based tomography is a powerful tool.
It enables assessment of collagen structural organization within the cornea.