Quantitative analysis of ring-shaped (crater-like) features at the tear film–epithelial interface of the rabbit cornea as assessed by scanning electron microscopy

Michael J. Doughty

Research output: Contribution to journalArticle

Abstract

Purpose: To develop a method for the quantitative analyses of the ring-shaped features usually observed on the surface of mammalian corneal epithelial cells. Methods: The corneas of 10 young adult female albino rabbits were prepared for scanning electron microscopy (SEM) by fixation in 2% glutaraldehyde in 80 mM cacodylate buffer (pH 7.2 to 7.4, 320 to 340 mOsm/kg) and single images taken at 1000× of the central region of the corneal epithelial surface. On enlarged prints at 5000× final magnification, cell-cell borders were marked as were the outlines of the ring-shaped features. Cell areas and the dimensions of the ring-shaped features were measured and the cell characteristics in terms of electron reflex (light, medium, and dark) as well as the number of apices (sides) also noted. Results: The ring-shaped features had an overall density of 2.5/100 µm, or an average of 6.5/cell. With the corneal epithelial cells being small and light in appearance (median area 36 µm), medium in size and appearance (median area 151 µ m), or larger and darker (median area 476 µ m), the number of features/cell was very strongly associated with both the cell type (p < 0.001) and the cell area (p < 0.001). The ring-shaped features could be between 0.65 and 5.5 µm in diameter, but sizes more than 3 µm were most unusual. Ring-shaped features of light cells were smallest (median diameter 1.35 µ m), slightly larger on medium cells (1.55 µm), and largest on the dark cells (1.90 µm). Conclusions: The ring-like features seen on SEM photomicrographs of rabbits (also known as craters or holes) are reproducible in terms of their overall density per unit surface area as well as their diameter. Such methods should be useful for objective analysis of these features after various treatments of the ocular surface.
Original languageEnglish
Pages (from-to)999-1010
Number of pages12
JournalCurrent Eye Research
Volume31
Issue number12
DOIs
Publication statusPublished - Dec 2006

Fingerprint

Tears
Electron Scanning Microscopy
Cornea
Rabbits
Epithelial Cells
Cacodylic Acid
Light
Glutaral
Reflex
Young Adult
Buffers
Cell Count
Electrons

Keywords

  • corneal epithelium
  • epithelial holes
  • ocular surfaces
  • rabbits

Cite this

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title = "Quantitative analysis of ring-shaped (crater-like) features at the tear film–epithelial interface of the rabbit cornea as assessed by scanning electron microscopy",
abstract = "Purpose: To develop a method for the quantitative analyses of the ring-shaped features usually observed on the surface of mammalian corneal epithelial cells. Methods: The corneas of 10 young adult female albino rabbits were prepared for scanning electron microscopy (SEM) by fixation in 2{\%} glutaraldehyde in 80 mM cacodylate buffer (pH 7.2 to 7.4, 320 to 340 mOsm/kg) and single images taken at 1000× of the central region of the corneal epithelial surface. On enlarged prints at 5000× final magnification, cell-cell borders were marked as were the outlines of the ring-shaped features. Cell areas and the dimensions of the ring-shaped features were measured and the cell characteristics in terms of electron reflex (light, medium, and dark) as well as the number of apices (sides) also noted. Results: The ring-shaped features had an overall density of 2.5/100 µm, or an average of 6.5/cell. With the corneal epithelial cells being small and light in appearance (median area 36 µm), medium in size and appearance (median area 151 µ m), or larger and darker (median area 476 µ m), the number of features/cell was very strongly associated with both the cell type (p < 0.001) and the cell area (p < 0.001). The ring-shaped features could be between 0.65 and 5.5 µm in diameter, but sizes more than 3 µm were most unusual. Ring-shaped features of light cells were smallest (median diameter 1.35 µ m), slightly larger on medium cells (1.55 µm), and largest on the dark cells (1.90 µm). Conclusions: The ring-like features seen on SEM photomicrographs of rabbits (also known as craters or holes) are reproducible in terms of their overall density per unit surface area as well as their diameter. Such methods should be useful for objective analysis of these features after various treatments of the ocular surface.",
keywords = "corneal epithelium, epithelial holes, ocular surfaces, rabbits",
author = "Doughty, {Michael J.}",
note = "Wrong title submitted by academic. Corrected TM 9-12-16",
year = "2006",
month = "12",
doi = "10.1080/02713680601001103",
language = "English",
volume = "31",
pages = "999--1010",
journal = "Current Eye Research",
issn = "0271-3683",
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TY - JOUR

T1 - Quantitative analysis of ring-shaped (crater-like) features at the tear film–epithelial interface of the rabbit cornea as assessed by scanning electron microscopy

AU - Doughty, Michael J.

N1 - Wrong title submitted by academic. Corrected TM 9-12-16

PY - 2006/12

Y1 - 2006/12

N2 - Purpose: To develop a method for the quantitative analyses of the ring-shaped features usually observed on the surface of mammalian corneal epithelial cells. Methods: The corneas of 10 young adult female albino rabbits were prepared for scanning electron microscopy (SEM) by fixation in 2% glutaraldehyde in 80 mM cacodylate buffer (pH 7.2 to 7.4, 320 to 340 mOsm/kg) and single images taken at 1000× of the central region of the corneal epithelial surface. On enlarged prints at 5000× final magnification, cell-cell borders were marked as were the outlines of the ring-shaped features. Cell areas and the dimensions of the ring-shaped features were measured and the cell characteristics in terms of electron reflex (light, medium, and dark) as well as the number of apices (sides) also noted. Results: The ring-shaped features had an overall density of 2.5/100 µm, or an average of 6.5/cell. With the corneal epithelial cells being small and light in appearance (median area 36 µm), medium in size and appearance (median area 151 µ m), or larger and darker (median area 476 µ m), the number of features/cell was very strongly associated with both the cell type (p < 0.001) and the cell area (p < 0.001). The ring-shaped features could be between 0.65 and 5.5 µm in diameter, but sizes more than 3 µm were most unusual. Ring-shaped features of light cells were smallest (median diameter 1.35 µ m), slightly larger on medium cells (1.55 µm), and largest on the dark cells (1.90 µm). Conclusions: The ring-like features seen on SEM photomicrographs of rabbits (also known as craters or holes) are reproducible in terms of their overall density per unit surface area as well as their diameter. Such methods should be useful for objective analysis of these features after various treatments of the ocular surface.

AB - Purpose: To develop a method for the quantitative analyses of the ring-shaped features usually observed on the surface of mammalian corneal epithelial cells. Methods: The corneas of 10 young adult female albino rabbits were prepared for scanning electron microscopy (SEM) by fixation in 2% glutaraldehyde in 80 mM cacodylate buffer (pH 7.2 to 7.4, 320 to 340 mOsm/kg) and single images taken at 1000× of the central region of the corneal epithelial surface. On enlarged prints at 5000× final magnification, cell-cell borders were marked as were the outlines of the ring-shaped features. Cell areas and the dimensions of the ring-shaped features were measured and the cell characteristics in terms of electron reflex (light, medium, and dark) as well as the number of apices (sides) also noted. Results: The ring-shaped features had an overall density of 2.5/100 µm, or an average of 6.5/cell. With the corneal epithelial cells being small and light in appearance (median area 36 µm), medium in size and appearance (median area 151 µ m), or larger and darker (median area 476 µ m), the number of features/cell was very strongly associated with both the cell type (p < 0.001) and the cell area (p < 0.001). The ring-shaped features could be between 0.65 and 5.5 µm in diameter, but sizes more than 3 µm were most unusual. Ring-shaped features of light cells were smallest (median diameter 1.35 µ m), slightly larger on medium cells (1.55 µm), and largest on the dark cells (1.90 µm). Conclusions: The ring-like features seen on SEM photomicrographs of rabbits (also known as craters or holes) are reproducible in terms of their overall density per unit surface area as well as their diameter. Such methods should be useful for objective analysis of these features after various treatments of the ocular surface.

KW - corneal epithelium

KW - epithelial holes

KW - ocular surfaces

KW - rabbits

U2 - 10.1080/02713680601001103

DO - 10.1080/02713680601001103

M3 - Article

VL - 31

SP - 999

EP - 1010

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 12

ER -