Electron microscopy of endothelial cells in culture: III. Freeze-fracturing

Summary Endothelial cells can be examined by freeze-fracturing and replication techniques performed on pellets of endothelial cells scraped off their culture substratum or, alternatively, cells grown on pieces of cover glass can be fractured as monolayers. Deep etching can be performed on either preparation provided that the cryoprotectant glycerol is omitted and water, which sublimes away during the etching process, is substituted.The cleaning and handling of the replicas for subsequent viewing in the electron microscope must be performed with care and are described in detail.