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A. Zhivov, O. Stachs, R. Guthoff; In vivo Three–Dimensional Confocal Laser Scanning Microscopy of Corneal Surface and Epithelium Under Various Clinical Conditions . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5010.
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Evaluation of in vivo visualization technique for corneal surface and epithelium characterization by means of 3–D confocal laser scanning microscopy
Ten human corneae (three from healthy volunteers, one with bullous keratopathy, 5 cases after penetraiting keroplasty and one with corneal erosion) was examined by an in–house developed Rostock Cornea Module in combination with commercial available confocal laser scanning system (Heidelberg Retina Tomograph II, Heidelberg Engineering GmbH, Germany). Raw data were converted using ImageJ (NIH, USA) for 3D–reconstruction. Surface characterization was done using AMIRA 3.1 (TGS Inc, USA) software package.
Three dimensional reconstruction was done and selective oblique sections were examined to demonstrate alterations of corneal surface and epithelium. In healthy volunteers it was found physiological surface defects originated from apoptotic superficial cells without contact with underlying wing cell. Bulluous keratopathy was characterized by irregular bullae at the level of superficial and wing cells and edema cells of all epithelial layers. In keratoplasty corneae the epitheliasation of donor–recipient crossover zone and suture epitheliasation could be seen. Reconstruction of a corneal erosion zone showed the defect extending down to Bowmans membrane and allowed the visualization of the dynamic of wound healing.
Three dimensional confocal laser scanning microscopy allows in vivo visualization and analysis of corneal surface and spatial arrangement of cells of corneal epithelium in normal and pathological cornea. The method provides a basis for further studies of alterations of the cellular arrangement and epithelial innervation in corneal diseases. Such studies may help knclarify the mechanism of turn over of epithelial cells as well as wound healing under various clinical and experimental conditions.
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