Selective insertion of lipid probes from the external milieu into raft regions of the cell membrane would enable targeted studies of raft molecular organization and of raft-associated peptide and glycan components. In this work, we compared the insertion of several synthetic glycolipids into raft and non-raft membrane areas of Raji and EA.hy 926 endothelial cells. A glyco-cholesterol derivative in which the 3β oxygen atom is replaced by NH was selected as a candidate raft-selective probe. Although this glycolipid also inserted into other membrane areas, its raft-to-non-raft distribution ratio was higher than that of its -analog (natural 3β--cholesterol).