Localization of Sphingomyelin in Cholesterol Domains by Imaging Mass Spectrometry

The location of each lipid in a palmitoyloleoylphosphatidylcholine/18:0 sphingomyelin/cholesterol monolayer system is laterally resolved using imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) without the necessity of adding fluorescent labels. This system of coexisting immiscible liquid phases shows cholesterol domains with sizes and shapes comparable to those in the fluorescence microscopy literature. The results show that SM localizes with cholesterol and that palmitoyloleoylphosphatidylcholine is excluded. Moreover, the segregation is not complete, and there is a small amount of both phospholipids distributed throughout.