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Martin Kronschlager, Nooshin TalebiZadeh, Zhaohua Yu, Per Söderberg, Gullstrand lab; A new experimental model for studies of capsule bound epithelial cells in the rat. Invest. Ophthalmol. Vis. Sci. 2013;54(15):471.
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© ARVO (1962-2015); The Authors (2016-present)
Lens epithelial cells are essential for keeping the lens transparent. We decided to develop an experimental model that allows studies of complete native lens epithelial cell populations attached to their anterior lens capsule. It should be also possible to transfer the capsule for incubation, thus allowing sequential imaging with various markers.
The experimental animal was the Sprague Dawley Rat. Using an operation microscope, the eyes were enucleated and the lenses were extracted from the back side of the eye. The extracted lens was atraumatically immersed in PBS in a Petri dish. Remnants of the ciliary body were removed without touching the lens poles. The lens was then transferred to a microscope slide containing a pool of PBS with the anterior face down. A capsulorhexis at the posterior capsule was performed with a needle. While extracting the nucleus by hydrodissection, a 0.2 mm Abulon Ultra fishing line, formed to a 3 mm diameter ring, was inserted in the capsule. Excessive cortex material was removed with a needle. Finally, fixation with 4 % paraformaldehyde and staining with DAPI was performed.
The fluorescence pictures showed capsule bound complete sheets of epithelial cells.
We have developed a new model for experimental studies of populations of capsule bound lens epithelial cells. The model allows sequential microscopical imaging, interrupted by tissue incubation The nylon ring provides easy handling of the lens capsule with the lens epithelial cells attached to a flat capsule.
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