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C-60-SIMS studies of glycerophospholipid in a LIPID MAPS model system: KDO2-Lipid A stimulated RAW 264.7 cells

M. K. Passarelli ; Andrew G Ewing (Institutionen för kemi och molekylärbiologi ; Institutionen för kemi- och bioteknik, Analytisk kemi) ; N. Winograd
Surface and Interface Analysis (0142-2421). Vol. 45 (2013), 1, p. 298-301.
[Artikel, refereegranskad vetenskaplig]

Although secondary ion mass spectrometry (SIMS) has been successfully employed for mapping lipid distributions at the cellular level, the identification of intact lipid species in situ is often complicated by isobaric interference. The high mass resolution and tandem MS capabilities of a C60-QSTAR hybrid instrument has been utilized to identify over 50 lipid species from mouse macrophages (RAW 264.7). In this investigation, lipid assignments made based on mass accuracy were confirmed with tandem MS analyses. Data obtained from C60-SIMS was compared to liquid chromatography (LC)-MS data obtained by the LIPID MAPS consortium. A majority of the lipids detected with LC-MS, but not detected with C60-SIMS, were present at concentrations below 2.0 pmol/µg of DNA. Matrix-related effects prevented the detection of lipids with the glycerophosphoethanolamine (PE) headgroup, glycerophosphoserine (PS) headgroup and lipids with polyunsaturated fatty acyl chains in the C60-SIMS analyses. Lipid distributions obtained from a lawn of RAW 264.7 cells stimulated with the endotoxin KDO2-Lipid A were also studied. The results obtained with C60-SIMS agreed with the established LC-MS data for the glycerophosphoinositol lipid class (PI) with adequate molecular sensitivity achieved with as few as 500 cells.

Nyckelord: lipids, C60-QSTAR, KDO2 Lipid A, RAW 264.7, tandem MS, ion mass-spectrometry, macrophages, sims

The authors would like to thank Professor Robert Murphy and Dr. Joseph Hankin at the University of Colorado for insight on lipid chemistry. The authors acknowledge the LIPID MAPS Consortium (GM 069338-07) for financial support and LC-MS data. Also, NW acknowledges additional financial support from the National Institutes of Health (2R01 EB002016-16) and the National Science Foundation (# CHE-0908226), and AGE acknowledges the European Research Council, the Knut and Alice Wallenberg Foundation and the Swedish Research Council (VR) for support.

Denna post skapades 2013-07-02. Senast ändrad 2013-07-10.
CPL Pubid: 179640


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Institutionen för kemi och molekylärbiologi (GU)
Institutionen för kemi- och bioteknik, Analytisk kemi (2006-2014)



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