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Browsing by Author "Hänninen, Oona"

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  • Hänninen, Oona (University of HelsinkiHelsingin yliopistoHelsingfors universitet, 2014)
    The morphology of a normal feline cornea and two sequestra were characterised using light microscopy (LM) and Transmission electron microscopy (TEM). A modern technique, serial block face scanning electron microscopy (SB-EM), was used to reconstruct a three-dimensional model of a feline corneal squamous epithelial cell and the apical corneal surface. Feline corneal sequestra are demarcated, pigmented plaques affecting the corneal stroma. The aetiology, pathogenesis and cause of the pigmentation remain elusive despite previous studies. TEM analysis has been applied in the study of sequestra, but further studies on sequestrum ultrastructure are needed to identify the cause of the pigmentation and reveal the role of keratocytes in the pathogenesis of the lesion. Three-dimensional techniques have not been used before in the study of feline corneal sequestra, so this served as a pilot study with the aim to modify a suitable protocol for preparing corneal specimens for SB-EM imaging. LM and TEM analysis of the control specimen concurred with previous findings of normal feline corneal anatomy. Normal stroma displayed organised lamellae, and keratocytes contained abundant rough endoplasmic reticulum associated with secretion of collagen filaments. The mild sequestrum illustrated detached stromal lamellae and apoptotic or necrotic keratocytes. The severe sequestrum depicted the regular appearance of stromal lamellae. Intact, though abnormal, keratocytes contained and were surrounded by numerous electron lucent profiles. The structures may have been pigment granules that washed away during specimen preparation. SB-EM analysis of feline corneal epithelial cell ultrastructure revealed intricate finger like protrusions which adjacent cells use to interlock with each other. The nucleus was discoid with a flat apical and a slightly convex posterior surface. Three different surface structures of apical squamous epithelial cells were identified: microvilli, labyrinthine microplicae, and a combination of these. A suitable protocol for the preparation of corneal specimens for SB-EM was achieved. This study can be continued in attempt to image corneal stroma and keratocytes. With additional control and sequestrum specimens the morphology of keratocytes in the normal feline cornea and corneal sequestra could be compared. Differences may indicate that keratocytes play a role in the pathogenesis of corneal sequestra.