97th DOG Annual Meeting 1999
ASSESSMENT OF KERATOCYTE ARCHITECTURE IN KERATOCONIC TREPHANATES BY CONFOCAL MIKROSKOPY
H.-H. Becker 1, M. Walke 1, D. G. Weiss ²
Recent studies of our group have presented data about confocal visualisation of keratocyte morphology and its 3-dimensional network in healthy, human cornea (CO). This investigation was set up to determine the stromal cell-architecture in keratoconic trephanates (KT) ex-vivo.
Methods: After perforating keratoplasty KT were instantly stained (Live/Dead Kit, Molecular Probes, USA). Vitality testing was performed in the corneal buttons followed by high resolution imaging performed by confocal laser scanning fluorescence microscopy (Nikon Diaphot 300/ Noran Odyssey XL). The digital processed stromal data were 3-dimensional reconstructed (VoxelView, software package) and quantitativly analysed (InterVision, software package).
Results: Morphometry of the posterior stroma in the region of maximal ectasia (KT, n = 7) revealed a significant reduction of volume density (p < 0,025) compared to healthy Co; cell density was reduced but not significantly. Comparing intact posterior Co with KT periphery no differences were detected.
Conclusions: We subdifferenciated the posterior stroma in human keratoconus. Further investigations might evaluate the role of the pathological keratocytes in the pathogenesis of keratoconus.
1) University-Eye-Hospital Rostock, Doberaner-Str. 140, D-18055 Rostock
2) Center of Microscopy Warnemünde, Institute of Cell Physiology, University of Rostock, Uniplatz 2, D-18055 Rostock