title: Automated tri-image analysis of stored corneal endothelium.
abstract: 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.
authors.affiliation: Ophthalmology Department, Bellevue Hospital, 25 Bd Pasteur, 42055 Saint-Etienne Cedex 2, France. philippe.gain@univ-st-etienne.fr
authors.forename: P
authors.initials: P
authors.lastname: Gain
authors.forename: G
authors.initials: G
authors.lastname: Thuret
authors.forename: L
authors.initials: L
authors.lastname: Kodjikian
authors.forename: Y
authors.initials: Y
authors.lastname: Gavet
authors.forename: P H
authors.initials: PH
authors.lastname: Turc
authors.forename: C
authors.initials: C
authors.lastname: Theillere
authors.forename: S
authors.initials: S
authors.lastname: Acquart
authors.forename: J C
authors.initials: JC
authors.lastname: Le Petit
authors.forename: J
authors.initials: J
authors.lastname: Maugery
authors.forename: L
authors.initials: L
authors.lastname: Campos
mesh_terms: D000368:Aged; D002452:Cell Count; D003315:Cornea; D004728:Endothelium, Corneal / Q000166:cytology*; D005125:Eye Banks; D006801:Humans; D007091:Image Processing, Computer-Assisted; D018715:Microscopy, Video; D008875:Middle Aged; D012680:Sensitivity and Specificity; D014021:Tissue Preservation