June 2015
Volume 56, Issue 7
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ARVO Annual Meeting Abstract  |   June 2015
Measurement of In Vivo Three-dimensional Corneal Cell Density and Size Using Two-photon Imaging in C57BL/6 Mice
Author Affiliations & Notes
  • Hongmin Zhang
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Siyu He
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Susu Liu
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Guoming Chen
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Junjie Zhang
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Shengtao Sun
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Liya Wang
    Henan Key Laboratory of Keratopathy, Henan Eye Institute Henan Eye Hospital, Zhengzhou, China
  • Footnotes
    Commercial Relationships Hongmin Zhang, None; Siyu He, None; Susu Liu, None; Guoming Chen, None; Junjie Zhang, None; Shengtao Sun, None; Liya Wang, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4900. doi:
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      Hongmin Zhang, Siyu He, Susu Liu, Guoming Chen, Junjie Zhang, Shengtao Sun, Liya Wang; Measurement of In Vivo Three-dimensional Corneal Cell Density and Size Using Two-photon Imaging in C57BL/6 Mice. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4900.

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

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Abstract

Purpose: To measure the cell size and cell density in five layers of the central cornea: the superficial epithelium, the basal epithelium, the anterior stroma, the posterior stroma, and the endothelium in the widely used inbred C57BL/6 mouse strain based on in vivo three-dimensional (3D) Two-photon (2PH) imaging

Methods: Corneas were scanned using a 2PH laser scanning fluorescence microscope after staining with plasma membrane stain and Hoechst 33342. Cell density and Cell size measurements including cell surface area, cell volume, nuclear surface area, and nuclear volume were automatically quantified using the IMARIS software. The cell and nuclear surface-area-to-volume ratio (S:V ratio) and the cell nuclear-cytoplasmic (N:C) ratio were calculated.

Results: The cell density was highest in the basal epithelium layer and lowest in the posterior stroma. The cell surface-area was highest in the anterior stroma, and the cell and nuclear volume was highest in the superficial epithelium. The cell surface-area, the cell and nuclear volume were both lowest in the basal epithelium. The cell and nuclear S:V ratio was highest in the basal epithelium and lowest in the superficial epithelium. The N:C ratio was highest in the basal epithelial cells and lowest in the posterior keratocytes.

Conclusions: The present study is the first to quantify the cell density and size parameters in the five layers of the central cornea. These data provide important cell morphology features for the study of corneal physiology, pathology and disease in C57BL/6 mice.

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