Repeatability of in vivo corneal confocal microscopy to quantify corneal nerve morphology

Cornea. 2013 May;32(5):e83-9. doi: 10.1097/ICO.0b013e3182749419.


Purpose: To establish intraobserver and interobserver repeatability, agreement, and symmetry of corneal nerve fiber (NF) morphology in healthy subjects using in vivo corneal confocal microscopy.

Methods: Nineteen subjects underwent in vivo corneal confocal microscopy (Heidelberg Retinal Tomograph III Rostock Cornea Module) at baseline and 7 days apart. Bland-Altman plots were generated to assess agreement, and the intraclass correlation coefficient and coefficient of repeatability were calculated to estimate intraobserver and interobserver repeatability for corneal NF density (numbers per square millimeter), nerve branch density (NBD; numbers per square millimeter), NF length (millimeters per square millimeter), and NF tortuosity coefficient. Symmetry between the right and left eyes was also assessed.

Results: Intraclass correlation coefficient and coefficient of repeatability for intraobserver repeatability were 0.66 to 0.74 and 0.17 to 0.64, for interobserver repeatability 0.54 to 0.93 and 0.15 to 0.85, and for symmetry 0.34 to 0.77 and 0.17 to 0.63, respectively. NBD demonstrated low repeatability.

Conclusions: This study demonstrates good repeatability for the manual assessment of all major corneal NF parameters with the exception of NBD, which highlights the difficulty in defining nerve branches and suggests the need for experienced observers or automated image analysis to ensure optimal repeatability.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Body Mass Index
  • Cholesterol / blood
  • Cornea / innervation*
  • Glycated Hemoglobin / metabolism
  • Humans
  • Microscopy, Confocal*
  • Observer Variation
  • Ophthalmic Nerve / anatomy & histology*
  • Reproducibility of Results
  • Young Adult


  • Glycated Hemoglobin A
  • hemoglobin A1c protein, human
  • Cholesterol