Time-of-flight secondary ion mass spectrometry (TOF-SIMS) enables chemically imaging the distributions of various lipid species in model membranes. However, discriminating the TOF-SIMS data of structurally similar lipids is very difficult because the high intensity, low mass fragment ions needed to achieve submicrometer lateral resolution are common to multiple lipid species. Here, we demonstrate that principal component analysis (PCA) can discriminate the TOF-SIMS spectra of four unlabeled saturated phosphatidylcholine species, 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero- 3-phosphocholine (DPPC), and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) according to variations in the intensities of their low mass fragment ions (m/z ≤ 200). PCA of TOF-SIMS images of phase-separated DSPC/DLPC and DPPC/DLPC membranes enabled visualizing the distributions of each phosphatidylcholine species with higher contrast and specificity than that of individual TOF-SIMS ion images. Comparison of the principal component (PC) scores images to atomic force microscopy (AFM) images acquired at the same membrane location before TOF-SIMS analysis confirmed that the PC scores images reveal the phase-separated membrane domains. The lipid composition within these domains was identified by projection of their TOF-SIMS spectra onto PC models developed using pure lipid standards. This approach may enable the identification and chemical imaging of structurally similar lipid species within more complex membranes.
ASJC Scopus subject areas
- Analytical Chemistry