The cholesterol/phospholipid and sphingomyelin/phospholipid ratios of red blood cells stored for 42 days were unchanged after storage. However, the total phospholipid concentration in the red cells decreased suggesting a loss of red cell membrane during storage. The 24 h survival of the stored red cells was assessed by the Cr technique in homologous donors. A decrease in fluorescence polarization of diphenylhexatriene incorporated into the membrane was observed following storage which correlated with survival. Thus, molecular defects in the lipid bilayer are associated with long-term storage of red blood cells.