freeze fracture

Method of specimen preparation for the electron microscope in which rapidly frozen tissue is cracked so as to produce a fracture plane through the specimen. The surface of the fracture plane is then shadowed by heavy metal vapour, strengthened by a carbon film, and the underlying specimen is digested away, leaving a replica that can be picked up on a grid and examined in the transmission electron microscope. The great advantage of the method is that the fracture plane tends to pass along the centre of lipid bilayers, and it is therefore possible to get en face views of membranes that reveal the pattern of integral membrane proteins. The E-face is the outer lamella of the plasma membrane viewed as if from within the cell, the P-face the inner lamella viewed from outside the cell. Fracture planes also often pass along lines of weakness such as the interface between cytoplasm and membrane, so that outer and inner membrane surfaces can be viewed. Further information about the structure can be revealed by freeze etching. Extremely rapid freezing followed by deep etching has allowed the structure of the cytoplasm to be studied without the artefacts that might be introduced by fixation.