Some modifications of freeze-fracture replica preparation

Freeze-fracture replicas are freed from lipid material by saponification in a hot sodium hydroxide solution and subsequently rinsed with 10% ethanol in distilled water. All manipulations are carried out with a glass bead fashioned onto a pasteur pipette. A stable, two-layered plastic support film has been developed to allow the use of grids with larger openings so that more of the specimen is observable.