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Updated: Jun 4, 2026
En Face Preparation of Mouse Blood Vessels
Published on: May 19, 2017
Endothelial glycocalyx structure in the intact carotid artery: a two-photon laser scanning microscopy study
Sietze Reitsma1, Mirjam G A oude Egbrink, Hans Vink
1Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, The Netherlands. s.reitsma@maastrichtuniversity.nl
Background:
The endothelial glycocalyx (EG) is the carbohydrate-rich luminal lining of endothelial cells that mediates permeability and blood cell-vessel wall interactions. To establish an atheroprotective role of the EG, adequate imaging and quantification of its properties in intact, viable, atherogenesis-prone arteries is needed.
Methods:
Carotid arteries of C57Bl6/J mice (n=22) were isolated including the bifurcation, mounted in a perfusion chamber, and perfused with fluorescent lectin wheat germ agglutinin-fluorescein isothiocyanate. The EG was visualized through the vessel wall using two-photon laser scanning microscopy. An image quantification protocol was developed to assess EG thickness, which was sensitive to hyaluronidase-induced changes.
Results:
In the lesion-protected common carotid artery, EG thickness was found to be 2.3 ± 0.1 μm (mean ± SEM), while the surface area devoid of (wheat germ agglutinin-sensitive) EG was 8.9 ± 4.2%. Data from the external carotid artery were similar (2.5 ± 0.1 μm; 9.1 ± 5.0%). In the atherogenesis-prone internal carotid artery the EG-devoid surface area was significantly higher (27.4 ± 5.5%, p<0.05); thickness at the remaining areas was 2.5 ± 0.1 μm.
Conclusion:
The EG can be adequately imaged and quantified using two-photon laser scanning microscopy in intact, viable mounted carotid arteries. Spatial EG differences could underlie atherogenesis.
