Imaging and quantification of endothelial cell loss in eye bank prepared DMEK grafts using trainable segmentation software

Curr Eye Res. 2014 Sep;39(9):894-901. doi: 10.3109/02713683.2014.887120. Epub 2014 Mar 3.


Purpose: To improve accuracy and efficiency in quantifying the endothelial cell loss (ECL) in eye bank preparation of corneal endothelial grafts.

Methods: Eight cadaveric corneas were subjected to Descemet Membrane Endothelial Keratoplasty (DMEK) preparation. The endothelial surfaces were stained with a viability stain, calcein AM dye (CAM) and then captured by a digital camera. The ECL rates were quantified in these images by three separate readers using trainable segmentation, a plug-in feature from the imaging software, Fiji. Images were also analyzed by Adobe Photoshop for comparison. Mean times required to process the images were measured between the two modalities.

Results: The mean ECL (with standard deviation) as analyzed by Fiji was 22.5% (6.5%) and Adobe was 18.7% (7.0%; p = 0.04). The mean time required to process the images through the two different imaging methods was 19.9 min (7.5) for Fiji and 23.4 min (12.9) for Adobe (p = 0.17).

Conclusions: Establishing an accurate, efficient and reproducible means of quantifying ECL in graft preparation and surgical techniques can provide insight to the safety, long-term potential of the graft tissues as well as provide a quality control measure for eye banks and surgeons. Trainable segmentation in Fiji software using CAM is a novel approach to measuring ECL that captured a statistically significantly higher percentage of ECL comparable to Adobe and was more accurate in standardized testing. Interestingly, ECL as determined using both methods in eye bank-prepared DMEK grafts exceeded 18% on average.

Keywords: Corneal transplant surgery; Descemet Membrane Endothelial Keratoplasty; endothelial cell loss; eye bank; trainable segmentation.

MeSH terms

  • Adult
  • Cell Count
  • Cell Survival
  • Corneal Endothelial Cell Loss / diagnosis*
  • Descemet Stripping Endothelial Keratoplasty*
  • Endothelium, Corneal / pathology*
  • Eye Banks*
  • Fluoresceins / metabolism
  • Fluorescent Dyes / metabolism
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Middle Aged
  • Software
  • Specimen Handling / methods*
  • Tissue Donors


  • Fluoresceins
  • Fluorescent Dyes
  • calcein AM