Basal cell carcinoma (BCC) is the most common skin malignancy. In fact, it is as common as the sum of all other skin malignancies combined and the incidence is rising. In this focused and histology-guided study, tissue from a patient diagnosed with aggressive BCC was analyzed by imaging mass spectrometry in order to probe the chemistry of the complex tumor environment. Time-of-flight secondary ion mass spectrometry using a (CO2)6 k+ gas cluster ion beam allowed a wide range of lipid species to be detected. Their distributions were then imaged in the tissue that contained small tumor islands that were histologically classified as more/less aggressive. Maximum autocorrelation factor (MAF) analysis highlighted chemical differences between the tumors and the surrounding stroma. A closer inspection of the distribution of individual ions, selected based on the MAF loadings, showed heterogeneity in signal between different microtumors, suggesting the potential of chemically grading the aggressiveness of each individual tumor island. Sphingomyelin lipids were found to be located in stroma containing inflammatory cells.

A set of basal cell carcinoma samples, removed by Mohs micrographic surgery and pathologically identified as having an aggressive subtype, have been analyzed using time-of-flight secondary ion mass spectrometry (SIMS). The SIMS analysis employed a gas cluster ion beam (GCIB) to increase the sensitivity of the technique for the detection of intact lipid species. The GCIB also allowed these intact molecular signals to be maintained while surface contamination and delocalized chemicals were removed from the upper tissue surface. Distinct mass spectral signals were detected from different regions of the tissue (epidermis, dermis, hair follicles, sebaceous glands, scar tissue, and cancerous tissue) allowing mass spectral pathology to be performed. The cancerous regions of the tissue showed a particular increase in sphingomyelin signals that were detected in both positive and negative ion mode along with increased specific phosphatidylserine and phosphatidylinositol signals observed in negative ion mode. Samples containing mixed more and less aggressive tumor regions showed increased phosphatidylcholine lipid content in the less aggressive areas similar to a punch biopsy sample of a nonaggressive nodular lesion.

Time-of-flight secondary ion mass spectrometry (ToF−SIMS) using a (CO 2 ) 6k + gas cluster ion beam (GCIB) was used to analyze Escherichia coli mutants previously identified as having impaired plasmid transfer capability related to the spread of antibiotic resistance. The subset of mutants selected were expected to result in changes in the bacterial envelope composition through the deletion of genes encoding for FabF, DapF, and Lpp, where the surface sensitivity of ToF−SIMS can be most useful. Analysis of arrays of spotted bacteria allowed changes in the lipid composition of the bacteria to be elucidated using multivariate analysis and confirmed through imaging of individual ion signals. Significant changes in chemical composition were observed, including a surprising loss of cyclopropanated fatty acids in the fabF mutant where FabF is associated with the elongation of FA(16:1) to FA(18:1) and not cyclopropane formation. The ability of the GCIB to generate increased higher mass signals from biological samples allowed intact lipid A (m/z 1796) to be detected on the bacteria and, despite a 40 keV impact energy, depth profiled through the bacterial envelope along with other high mass ions including species at m/z 1820 and 2428, attributed to ECA CYC , that were only observed below the surface of the bacteria and were notably absent in the depth profile of the lpp mutant. The analysis provides new insights into the action of the specific pathways targeted in this study and paves the way for whole new avenues for the characterization of intact molecules within the bacterial envelope.

This work assesses the potential of new water cluster-based ion beams for improving the capabilities of secondary ion mass spectrometry (SIMS) for in situ lipidomics. The effect of water clusters was compared to carbon dioxide clusters, along with the effect of using pure water clusters compared to mixed water and carbon dioxide clusters. A signal increase was found when using pure water clusters. However, when analyzing cells, a more substantial signal increase was found in positive ion mode when the water clusters also contained carbon dioxide, suggesting that additional reactions are in play. The effects of using a water primary ion beam on a more complex sample were investigated by analyzing brain tissue from an Alzheimer’s disease transgenic mouse model. The results indicate that the ToF-SIMS results are approaching those from MALDI as ToF-SIMS was able to image lyso-phosphocholine (LPC) lipids, a lipid class that for a long time has eluded detection during SIMS analyses. Gangliosides, sulfatides, and cholesterol were also imaged. Graphical abstract

Cholesteryl phytanate and triglycerides containing phytanic acid were separated from their normal fatty acid analogs by thin-layer chromatography. The presence of the branched-chain fatty acid makes the lipid less polar, and this effect becomes more pronounced as the number of phytanic acid residues in the triglycerides is increased. Phytanic acid was prepared from commercial phytol by catalytic hydrogenation, followed by catalytic oxidation. It contained 5% pristanic acid.