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A. Bernal, M. Lynn, P. Joshi, S. Decker, W. Lee, J.–M. Parel; Anatomical Study of the Lens Accommodative Apparatus Using Environmental–Scanning Electron Microscopy (E–SEM) . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1981.
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© ARVO (1962-2015); The Authors (2016-present)
To demonstrate the feasibility of using Environmental Scanning Electron Microscopy (E–SEM) to study the anatomy of the accommodation apparatus in hydrated, unfixed, uncoated cadaver lenses during simulated accommodation.
A custom–made, 8–segment miniature manual lens stretcher was designed to mount human cadaver lenses with the zonules, ciliary body and sclera attached, inside an ESEM (Philips XL30 ESEM–FEG). The lens stretcher allows imaging of the accommodative apparatus in the unstretched and stretched states. One fresh 82 y/o human eye was mounted in the stretcher, dissected to expose lens, zonules and ciliary body, and stretched 3mm in diameter. The tissue was allowed to stand at room temperature for 20 minutes before scanning. The scanning sequence was adjusted to produce a gradual dehydration into a "hydration window" in which the tissue is hydrated but all the surface moisture is removed allowing for very detailed pictures of the specimen. Images of the accommodation apparatus were acquired at low magnification (39–50X).
Sharp, high contrast, high depth of field images of the lens and its accommodative apparatus were obtained. Posterior zonules embedding in the hyaloid membrane were clearly seen with this technique, providing detail difficult to obtain with regular SEM due to the deleterious effects of fixation and dehydration.
This study demonstrates that ESEM can be used to provide new, valuable morphological information of the unfixed and hydrated accommodative apparatus in cadaver lenses.
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