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Automated tri-image analysis of stored corneal endothelium
  1. P Gain1,2,3,4,
  2. G Thuret1,2,3,4,
  3. L Kodjikian1,
  4. Y Gavet2,
  5. P H Turc2,
  6. C Theillere3,
  7. S Acquart3,
  8. J C Le Petit3,
  9. J Maugery1,
  10. L Campos4
  1. 1Ophthalmology Department, Bellevue Hospital, 25 Bd Pasteur, 42055 Saint-Etienne Cedex 2, France
  2. 2Ecole Nationale Supérieure des Mines de Saint-Etienne, 42000 Saint-Etienne, France
  3. 3French Blood Center, Cornea Bank of St-Etienne (Loire/Auvergne region), 25 Bd Pasteur, 42055 Saint-Etienne Cedex 2, France
  4. 4“Cell Death and Neoplasia” Research Group (EA 3063), Faculty of Medicine, 15 rue Ambroise Paré, 42023 Saint-Etienne Cedex, France
  1. Correspondence to: Professor Philippe Gain, Service d'Ophtalmologie (pavillon 50A), CHRU de Bellevue, 25 Boulevard Pasteur, 42055 Saint-Etienne Cedex 2, France; philippe.gain{at}


Background: Endothelial examination of organ culture stored corneas is usually done manually and on several mosaic zones. Some banks use an image analyser that takes account of only one zone. This method is restricted by image quality, and may be inaccurate if endothelial cell density (ECD) within the mosaic is not homogeneous. The authors have developed an analyser that has tools for automatic error detection and correction, and can measure ECD and perform morphometry on multiple zones of three images of the endothelial mosaic.

Methods: 60 human corneas were divided into two equal groups: group 1 with homogeneous mosaics, group 2 with heterogeneous ones. Three standard microscopy video images of the endothelium, graded by quality, were analysed either in isolation (so called mono-image analysis) or simultaneously (so called tri-image analysis), with 50 or 300 endothelial cells (ECs) counted. The automated analysis was compared with the manual analysis, which concerned 10 non-adjacent zones and about 300 cells. For each analysis method, failures and durations were studied according to image quality.

Results: All corneas were able to undergo analysis, in about 2 or 7.5 minutes for 50 and 300 ECs respectively. The tri-image analysis did not increase analysis time and never failed, even with mediocre images. The tri-image analysis of 300 ECs was always most highly correlated with the manual count, particularly in the heterogeneous cornea group (r=0.94, p<0.001) and prevented serious count errors.

Conclusions: This analyser allows reliable and rapid analysis of ECD, even for heterogeneous endothelia mosaics and mediocre images.

  • endothelium
  • cornea
  • organ culture
  • automatic analysis
  • multi-image analysis
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