Abstract
Purpose :
High resolution visualization of the eyeball is critical and essential for our understanding and management of eye diseases. In this study, we used tissue clearing and three dimensional imaging technology to achieve this goal and imaged a variety of ocular structures and cell types from both normal and disease conditions.
Methods :
Wildtype and transgenic mice of Prox-1-GFP, GFAP-GFP, or Thy1-YFP were used in the study. Eyeballs were harvested from normal or disease models of inflammation and glaucoma. Samples were infused with hydrogel monomers and heated for polymerization. Lipids were removed by electrophoresis. The transparent tissue-hydrogel hybrids with immunolabeling or endogenous fluorescence were imaged by a Zeiss light sheet microscope.
Results :
Optical transparency was achieved in the intact eyeballs from both normal and disease conditions. High resolution and three dimensional images and videos were obtained for a wide array of structures and cell types explored in this study from the anterior to the posterior segments, such as blood vessels, lymphatic vessels, corneal nerves, Schlemm’s canal, optic nerves, and astrocytes.
Conclusions :
To our knowledge, this study provides the first and comprehensive imaging of the intact eyeball using the tissue clearing and light sheet microscopic methods. Given that the eye is the window of the body, we anticipate this advanced technology will facilitate diverse applications in biomedical research inside and outside the eye.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.