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Morphologie. 2017 Jun;101(333):55-63. doi: 10.1016/j.morpho.2017.04.001. Epub 2017 May 12.

Electron microscopy approach for the visualization of the epithelial and endothelial glycocalyx.

Author information

1
Normandie University, Unirouen, INSA Rouen, CNRS, GPM-UMR6634, 76000 Rouen, France. Electronic address: laurence.chevalier@univ-rouen.fr.
2
Inserm, U1096, Normandie University, Unirouen, 76000 Rouen, France.
3
Department of Pathology, Rouen University Hospital, 76000 Rouen, France.
4
Inserm, U1096, Normandie University, Unirouen, 76000 Rouen, France; Department of Thoracy Surgery, Rouen University Hospital , 76000 Rouen, France.
5
Normandie University, Unirouen, INSA Rouen, CNRS, GPM-UMR6634, 76000 Rouen, France.

Abstract

This study presents a methodological approach for the visualization of the glycocalyx by electron microscopy. The glycocalyx is a three dimensional network mainly composed of glycolipids, glycoproteins and proteoglycans associated with the plasma membrane. Since less than a decade, the epithelial and endothelial glycocalyx proved to play an important role in physiology and pathology, increasing its research interest especially in vascular functions. Therefore, visualization of the glycocalyx requires reliable techniques and its preservation remains challenging due to its fragile and dynamic organization, which is highly sensitive to the different process steps for electron microscopy sampling. In this study, chemical fixation was performed by perfusion as a good alternative to conventional fixation. Additional lanthanum nitrate in the fixative enhances staining of the glycocalyx in transmission electron microscopy bright field and improves its visualization by detecting the elastic scattered electrons, thus providing a chemical contrast.

KEYWORDS:

Chemical contrast; Contraste chimique; Electron scattering; Glycocalyx; Lanthane; Lanthanum; Microscopie électronique en transmission; Perfusion; Perfusion fixation; Transmission Electron Microscopy

PMID:
28506708
DOI:
10.1016/j.morpho.2017.04.001
[Indexed for MEDLINE]

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