Cold acclimation is a complex adaptive mechanism by which plants survive freezing in winter. During cold acclimation, diverse intracellular and extracellular changes occur. Although most of these changes are related to the acquirement of freezing tolerance, the exact role of these changes in the attainment of freezing tolerance is still unclear. In this review, we suggest the possible role of some of these cold acclimation-induced changes in relation with increased freezing tolerance from the viewpoint of inhibition of freezing injury produced by close approach of membranes.