Three-dimensional confocal microscopy of the living cornea and ocular lens

Barry R. Masters

doi:10.1117/12.44304

1 July 1991 Three-dimensional confocal microscopy of the living cornea and ocular lens

Barry R. Masters

Proceedings Volume 1450, Biomedical Image Processing II; (1991) https://doi.org/10.1117/12.44304
Event: Electronic Imaging '91, 1991, San Jose, CA, United States

Abstract

The three-dimensional reconstruction of the optic zone of the cornea and the ocular crystalline lens has been accomplished using confocal microscopy and volume rendering computer techniques. A laser scanning confocal microscope was used in the reflected light mode to obtain the two-dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with a 488 nm wavelength. The microscope objective was a Leitz X25, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133 three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their 'beaded' cell borders, basal lamina, nerve plexus, nerve fibers, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in- situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers. The three-dimensional data sets of the cornea and the ocular lens were reconstructed in the computer using volume rendering techniques. Stereo pairs were also created of the two- dimensional ocular images for visualization. The stack of two-dimensional images was reconstructed into a three-dimensional object using volume rendering techniques. This demonstration of the three-dimensional visualization of the intact, enucleated eye provides an important step toward quantitative three-dimensional morphometry of the eye. The important aspects of three-dimensional reconstruction are discussed.

Citation Download Citation

Barry R. Masters "Three-dimensional confocal microscopy of the living cornea and ocular lens", Proc. SPIE 1450, Biomedical Image Processing II, (1 July 1991); https://doi.org/10.1117/12.44304

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