title: In vivo confocal microscopy of the corneal endothelium: comparison of three morphometry methods after corneal transplantation.
abstract: OBJECTIVE
  The purpose of this study was to assess the endothelium of corneal grafts by in vivo confocal microscopy (IVCM), and to evaluate an automated endothelial software system in comparison with a manual cell count and planimetry.
  
  
  METHODS
  Overall, 40 corneal grafts (20 deep anterior lamellar keratoplasties (DALKs) and 20 penetrating keratoplasties (PKs)) were assessed by scanning-slit IVCM. The endothelial cell density (ECD) was estimated with the automated and the manual cell count method of the instrument's Nidek Advanced Vision Information System (NAVIS) software. The results were compared with planimetry as the reference method, and the agreement was assessed.
  
  
  RESULTS
  The mean (±SD) automated ECD was 2278±524 cells/mm(2) (range 1167-3192 cells/mm(2)), whereas the manual cell count method gave significantly lower ECDs with a mean of 1213±677 cells/mm(2) (range 218-2440 cells/mm(2); P<0.001). The manual cell counts were also significantly lower than those by planimetry, with a mean ECD of 1617±813 cells/mm(2) (range 336-2941, P<0.001). Bland-Altman analyses indicated that the limits of agreement (LoA) between the automated and the planimetry method were -671 and +1992 cells/mm(2), whereas they were -1000 and +202 cells/mm(2) when comparing the manual cell counts with planimetry.
  
  
  CONCLUSIONS
  Following keratoplasty, the NAVIS automated method is likely to overestimate endothelial cell counts due to oversegmenting of the cell domains. Automated ECDs are substantially higher than those by the manual counting method or planimetry. The differences are considerably larger post-keratoplasty than for normal corneas, and the methods should not be used interchangeably.
authors.affiliation: Tennent Institute of Ophthalmology, Gartnavel General Hospital, Glasgow, UK. sven.jonuscheit@gcu.ac.uk
authors.forename: S
authors.initials: S
authors.lastname: Jonuscheit
authors.forename: M J
authors.initials: MJ
authors.lastname: Doughty
authors.forename: K
authors.initials: K
authors.lastname: Ramaesh
mesh_terms: D000328:Adult; D002452:Cell Count / Q000379:methods; D016039:Corneal Transplantation; D003430:Cross-Sectional Studies; D004728:Endothelium, Corneal / Q000166:cytology*; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods*; D008875:Middle Aged; D009885:Ophthalmology / Q000379:methods