Quantitative assessment of anteroposterior keratocyte density in the normal rabbit cornea

W. M. Petroll, K. Boettcher, P. Barry, D. Cavanagh, J. V. Jester

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Abstract

The anteroposterior keratocyte density distribution in the rabbit cornea was measured. Unsectioned tissue blocks from the central cornea of five rabbits were stained with propidium iodide and imaged using a Leica laser scanning confocal microscope. A z-series of images was acquired in each sample, from anterior to posterior stroma in either 3- or 8-μm steps. Software was developed to allow interactive marking of the keratocyte nuclei within each section of the z-series and for calculating cell density. For convenience, cell density was expressed as the number of cells per corneal volume element (CVE), where CVE is a newly defined volume unit with x, y, and z dimensions of 250, 250, and 10 μm, respectively. The calculated keratocyte density was 20.2 ± 1.0 cells/CVE (n = 5), which is equivalent to 32,360 ± 1,660 cells/mm3. The greatest density was underneath the epithelium (26.3 ± 2.5 cells/ CVE), the density then decreased linearly with depth to 15.2 ± 1.4 cells/CVE; there was a slight increase in density pre-Descemets membrane to 18.5 ± 3.5 cells/CVE. A 30% decrease in cell density over the entire anteroposterior stromal thickness was observed. To facilitate statistical analysis, the cell density was averaged over 5% thickness intervals from anterior to posterior cornea. A significant difference in mean cell density of these intervals was found (ANOVA, n = 20, p < 0.01). To further assess the density distribution, linear regression analysis was performed. A significant correlation was found between keratocyte density and stromal depth (R = - 0.94, n = 20, p < 0.05). We conclude that in the rabbit, keratocyte cell density is maximal underlying the epithelium and progressively decreases from anterior to posterior cornea.

Original languageEnglish (US)
Pages (from-to)3-9
Number of pages7
JournalCornea
Volume14
Issue number1
Publication statusPublished - 1995

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Keywords

  • Confocal microscopy
  • Corneal keratocyte
  • Three-dimensional imaging

ASJC Scopus subject areas

  • Ophthalmology

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