When Escherichia coli is frozen rapidly in saline and thawed slowly, survival is very low; however, the inclusion of 3% glycerol or 1% Tween 80 in the saline freezing menstruum results in near complete survival. The release of material from, and penetration of, substances into the cell indicate that both membrane and wall damage occur during freezing and thawing. Glycerol, under these conditions, is able to reduce severely both the damage to the wall and membrane, whereas Tween 80 prevents only membrane damage. This indicates that freezing and thawing in saline results in membrane damage which is lethal to the cell whereas wall damage which occurs is not detrimental to cell survival.