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U.C. Birk, J. von Eicken, O. Denk; Digitalized Evaluation of Endothelial Cells on Donor Corneas Using a Notebook-based Video System . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4709.
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Purpose: Development and validation of a new approach to a digitalised endothelial cell evaluation of donor corneas using a notebook-based video system. Methods: Endothelial cells of ten porcine cornea were examined immediately after excision of the corneoscleral segment. After swelling of cell borders with hypotonic BSS solution, endothelial cells were visualized by a Nikon TS 100 phase contract microscope, detected by an analogue high-resolution video camera (BC-71), digitalized by an ADVC-100 converter and analyzed in a Sony VAIO Notebook using commercially available image analysis software. To validate the quality of endothelial cell evaluation, we checked the number of vital endothelial cells after trypan blue and alizarin red staining by light microscopy. All counts were performed on endothelial cells of 10 porcine corneas in triplicate. Results: Mean endothelial density of porcine corneas was determined to be 3597 ± 370 cells/mm2 when evaluated by phase contrast microscopy after swelling of cell borders with hypotonic BSS and 3913 ± 301 cells/mm2 when evaluated by vital staining using trypan blue and alizarin red. The difference was statistically significant when the data were analyzed with student's t-test (p=0.04). When cell counting of individual corneas was compared, in two cases cell counting performed by phase contrast microscopy resulted in higher endothelial cell densities when compared to cell counting after vital staining. Conclusions: Endothelial cell counting using phase contrast microscopy and a notebook-based video system resulted in lower endothelial cell densities when compared to cell counting after vital staining on average. However, since the risk to overestimate endothelial cell density with phase contrast microscopy is small, the notebook-based video system used in the present study is deemed to be valid enough to be used for routine cell counting in a cornea bank.
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