Application of freeze-etch replication for the observation of the cell-extracellular matrix interface of cultured cells.
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.
Sawada
H
H
Department of Fine Morphology, University of Tokyo, Japan.
D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure*; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005613:Freeze Etching / Q000379:methods; D008854:Microscopy, Electron; D013048:Specimen Handling