The first quantitative insight has been obtained into the effects that volatile anesthetics have on the interactions and lateral organization of lipids in model membranes that mimic "lipid rafts". Specifically, nearest-neighbor recogntion measurements, in combination with Monte Carlo simulations, have been used to investigate the action of isoflurane, halothane, and chloroform on the compactness and lateral organization of cholesterol-rich bilayers of 1,2-dipalmitoylsn-glycero-3-phosphocholine (DPPC) in the liquid-ordered (l(o)) phase. All three anesthetics induce a similar weakening of sterol-phospholipid association, corresponding to ca. 30 cal/mol of lipid at clinically relevant concentrations. Monte Carlo lattice simulations show that the lateral organization of the l(o) phase, under such conditions, remains virtually unchanged. In sharp contrast to their action on the l(o) phase, these anesthetics have been found to have a similar strengthening effect on sterol-phospholipid association in the liquid-disordered (l(d)) phase. The possibility of discrete complexes being formed between DPPC and these anesthetics and the biological relevance of these findings are discussed.