Visualization of plasma membrane compartmentalization with patterned lipid bilayers

Micrometer-size patterned lipid bilayers containing liganded lipids are used to control the location and size of receptor clusters and enable direct visualization of structural reorganization of cellular components. Subsequent to concentration of Fcε receptor I, the mast cell receptor for IgE, and colocalized tyrosine phosphorylation activity, Lyn kinase and other proteins anchored to the inner leaflet of the plasma membrane redistribute selectively with the receptor clusters in a process that depends on actin polymerization. Surprisingly, outer leaflet components characteristically associated with lipid rafts do not detectably coredistribute with these inner leaflet components. Cell activation using patterned surfaces provides unique insights into cell membrane structural organization, revealing dynamic, large-scale uncoupling of inner and outer leaflet components of lipid rafts.