X‑ray Absorption Spectroscopy and Spectromicroscopy of Supported Lipid Bilayers

The C 1s, O 1s, and N 1s X-ray absorption spectra of three lipid species, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and 1,2-dioleoyl-3-trimethylammonium-propane (chloride salt) (DOTAP), have been recorded using transmission detection in a scanning transmission X-ray microscope. The spectra are presented on an absolute intensity scale (i.e., optical density per nm) to allow their use as reference standards for spectromicroscopic analysis of supported lipid bilayers. Examples of C 1s based spectromicroscopic mapping of saturated and unsaturated domains in dry lipid bilayers of DOPC and DSPC at several compositions are presented. The results are compared with fluorescence microscopy of the same area. Challenges for extending this work to studies of wet lipid bilayers interacting with antimicrobial peptides are discussed.