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Miltiadis K. Tsilimbaris, Eric Lesniewska, Stella Lydataki, Christian Le Grimellec, Jean P. Goudonnet, Ioannis G. Pallikaris; The Use of Atomic Force Microscopy for the Observation of Corneal Epithelium Surface. Invest. Ophthalmol. Vis. Sci. 2000;41(3):680-686.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To evaluate the feasibility of imaging normal corneal epithelium by
means of atomic force microscopy (AFM).
methods. Twelve normal corneas from six albino rabbits were examined using a
commercial atomic force microscope. Six corneas were examined in
balanced salt solution after fixation in glutaraldehyde 2.5% and six
without any fixation. Rectangular silicon nitride cantilevers with a
spring constant of 10 to 20 mN/m were used. The measured forces after
imaging were less than 100 pN. All reported images were made with
512 × 512-pixel definition with typical scan rates ranging from 1
to 5 Hz.
results. High-quality images of corneal epithelium surface were obtained from
fixed and unfixed specimens in magnifications ranging from ×2000 to×
2,000,000. Imaging of fixed specimens was always easier. In unfixed
specimens fuzzy images were very common, probably because of the
presence of the cell glycocalyx. AFM revealed the typical polygonal
corneal epithelial cells. The cell surface was covered by
microprojections; at cell borders the microprojections were arranged in
two characteristic parallel rows. Craterlike formations were revealed
in several specimens. The microprojections’ morphology and their
surface details were revealed using magnifications up to ×2,000,000.
Three-dimensional representation of the images facilitated better
understanding of the surface topography. Measurements in horizontal and
vertical plane were made using the section analysis tool.
conclusions. In this work the AFM parameters appropriate for corneal epithelium
imaging in physiological medium were defined. AFM represents a new
powerful tool for corneal epithelium imaging, and its application in
this field warrants further investigation.
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