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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)
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
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.
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.
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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