June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
A new experimental model for studies of capsule bound epithelial cells in the rat
Author Affiliations & Notes
  • Martin Kronschlager
    Neuroscience/Ophthalmology, University of Uppsala, Uppsala, Sweden
  • Nooshin TalebiZadeh
    Neuroscience/Ophthalmology, University of Uppsala, Uppsala, Sweden
  • Zhaohua Yu
    Neuroscience/Ophthalmology, University of Uppsala, Uppsala, Sweden
  • Per Söderberg
    Neuroscience/Ophthalmology, University of Uppsala, Uppsala, Sweden
  • Footnotes
    Commercial Relationships Martin Kronschlager, None; Nooshin TalebiZadeh, None; Zhaohua Yu, None; Per Söderberg, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 471. doi:
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    • Get Citation

      Martin Kronschlager, Nooshin TalebiZadeh, Zhaohua Yu, Per Söderberg, ; 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)

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Abstract

Purpose: 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.

Methods: 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.

Results: The fluorescence pictures showed capsule bound complete sheets of epithelial cells.

Conclusions: 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.

Keywords: 445 cataract • 554 immunohistochemistry  
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