Image processing techniques in a preliminary morphometric characterization of corneal epithelium by scanning electron microscopy

Several processing techniques of digital images allowed us to quantify the percentage of cell surface covered by microprojections (microvilli or microplicae) (SCM), the adhesion between epithelial cells by the parameter intercellular junctions (IJ), the size (cell area), shape (cell shape) and shade (cell shade) of cells on the corneal epithelium of nine rabbits. The data were analyzed and the epithelial cells were classified into three groups by cluster analysis. Assuming the representativeness of the sample, our findings suggest that for a normal corneal epithelium, 80% of the cells could show SCM >41%, and IJ >0.98 (being one a cell to cell junction without disruptions). Standard deviations of cell shade lower than 21 gray levels could indicate a tendency to lose the cell shade mosaic. Normal corneas could show a majority of cells (54–69%) included in group 2 with smaller mean size (80% of cells with cell area <242 μm²), higher SCM (80% of cells with SCM >44.83%), polygonal mean shape and brighter shade. Group 1 (15–30% of cells) could show a larger mean size (80% of cells with cell area >494 μm²), lower SCM (although 80% of cells with SCM >32.61%), circular mean shape and darker electron reflex. Group 3 could display a medium mean size, higher SCM (similar to group 2), circular mean shape (similar to group 1), and brighter shade. These analyses could possibly be used to further assess the corneal response to ocular drugs, contact lens, and surgical procedures or to discriminate between pathology stages. Microsc. Res. Tech. 73:1059–1066, 2010. © 2010 Wiley‐Liss, Inc.