June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Overestimation of Corneal Endothelial Cell Density by Selecting Smaller Areas to Count the Cells in Images of In Vivo Confocal Microscopy
Author Affiliations & Notes
  • Ahmad Kheirkhah
    Ophthalmology, Massachusetts Eye and Ear Infirmary, Boston, MA
  • Ujwala Saboo
    Ophthalmology, Massachusetts Eye and Ear Infirmary, Boston, MA
  • Reza Dana
    Ophthalmology, Massachusetts Eye and Ear Infirmary, Boston, MA
  • Footnotes
    Commercial Relationships Ahmad Kheirkhah, None; Ujwala Saboo, None; Reza Dana, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1148. doi:
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      Ahmad Kheirkhah, Ujwala Saboo, Reza Dana; Overestimation of Corneal Endothelial Cell Density by Selecting Smaller Areas to Count the Cells in Images of In Vivo Confocal Microscopy. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1148.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Laser scanning in vivo confocal microscopy (LS-IVCM) is commonly used to measure corneal endothelial cell density (CECD). However, due to corneal applanation during imaging, it is common to see dark non-focused areas in LS-IVCM images which make it impossible to count all corneal endothelial cells in the image. It is unclear whether counting endothelial cells in smaller areas in LS-IVCM images will affect the measured CECD.

Methods: 32 endothelial images by LS-IVCM (Heidelberg Retina Tomograph 3 with Rostock Corneal Module; Heidelberg, Germany) with different endothelial cell densities were analyzed. In each image (160 000 µm2), endothelial cells were counted within the following areas: 80 000, 40 000, 20 000, 10 000, 5 000, and 2 500 µm2. The cell numbers counted in these areas were then expressed as cell/mm2, and the CECD were compared.

Results: The mean measured CECD was 2007 ± 750 cells/mm2, 2109 ± 764 cells/mm2, 2149 ± 770 cells/mm2, 2290 ± 750 cells/mm2, 2495 ± 758 cells/mm2, and 2760 ± 854 cells/mm2 for areas of 80 000 µm2, 40 000 µm2, 20 000 µm2, 10 000 µm2, 5 000 µm2, and 2 500 µm2, respectively; the differences between all these groups were statistically significant (all P<0.05). Compared to the CECD in 80 000-µm2 area, the mean percentage of overestimation was 6.2 ± 6.7% for the 40 000-µm2 area, 8.9 ± 11.4% for the 20 000-µm2 area, 18.7 ± 21.1% for the 10 000-µm2 area, 27.9 ± 23.6% for the 5 000-µm2 area, and 45.6 ± 34.9% for the 2 500-µm2 area. There were significant negative correlations between the measured CECD for the 80 000-µm2 area and the percentage of overestimation in the smaller areas (R= -0.44 to -0.71, all P<0.05).

Conclusions: Counting endothelial cells in smaller areas of LS-IVCM images results in a significant overestimation of CECD.

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