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Application of freeze-etch replication for the observation of the cell-extracellular matrix interface of cultured cells
Authors:Hajime Sawada
Abstract:In order to investigate the ultrastructural three-dimensional relationship between extracellular matrices (ECM) and the plasma membranes of cultured cells, a freeze-etch replica method was devised. Bovine corneal endothelial cells were cultured on a Collodion film which covered a hole punched in a plastic coverslip, and were quickly frozen with a slammer with their basal surface facing a liquid nitrogen-cooled copper block. The cells were placed upside-down in a Balzers freeze-fracture machine and freeze-etched, and then platinum-carbon replicas were obtained. The structure of the ECM-plasma membrane interface was observed successfully and so this technique provides a new approach for investigating the ECM-plasma membrane (matrix-receptor) relationship.
Keywords:Cultured cell  quick-freezing  freeze-etching  extracellular matrix  plasma membrane
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