In situ lipidomics of Staphylococcus aureus osteomyelitis using imaging mass spectrometry

Osteomyelitis occurs when Staphylococcus aureus invades the bone microenvironment, resulting in a bone marrow abscess with a spatially defined architecture of cells and biomolecules. Imaging mass spectrometry and microscopy are invaluable tools that can be employed to interrogate the lipidome of S. aureus-infected murine femurs to reveal metabolic and signaling consequences of infection. Here, nearly 250 lipids were spatially mapped to healthy and infection-associated morphological features throughout the femur, establishing composition profiles for tissue types. Ether lipids and arachidonoyl lipids were significantly altered between cells and tissue structures in abscesses, suggesting their roles in abscess formation and inflammatory signaling. Sterols, triglycerides, bis(monoacylglycero)phosphates, and gangliosides possessed ring-like distributions throughout the abscess, suggesting a hypothesized dysregulation of lipid metabolism in a population of cells that cannot be discerned with..., All MALDI IMS data were acquired using a timsTOF fleX (Bruker Daltonics, Bremen, Germany).88 All imaging experiments were operated in qTOF mode with TIMS deactivated unless otherwise noted. For acquisition of the main dataset, a pitch offset strategy was employed. The SmartBeam 3D 10 kHz frequency tripled Nd:YAG laser (355 nm) was focused and beam scan activated to produce a 10 μm x 10 μm burn pattern. The z position of the stage was adjusted to keep the laser focused and account for additional height of the mount and Cryofilm. Positive ion data were acquired first with a stage pitch of 20 μm. Stage coordinates in flexImaging (Bruker Daltonics) were then offset 10 μm in each x-y dimension, and negative ion data were acquired with the same 20 μm stage pitch. This strategy yields an effective spatial resolution of 20 μm for both polarities from the same pixel regions. For high spatial resolution (10 μm) experiments, beam scan was still implemented, but the stage pitch was set to 10 μm. Th..., , # Data from: In situ lipidomics of Staphylococcus aureus osteomyelitis using imaging mass spectrometry

[https://doi.org/10.5061/dryad.bnzs7h4jz](https://doi.org/10.5061/dryad.bnzs7h4jz)

Included are all imaging mass spectrometry datasets used for the main figures of this manuscript

Also included is a spreadsheet entitled \"PCCValues(Figure7)\" which gives the raw PCC values from Figure 7.

## Description of the data and file structure

All data are in vendor-neutral imzml format, which is common for imaging mass spectrometry datasets. A cohort of three infected mice and three mock-infected mice were used for the main dataset involved in the majority of experimental analyses (**Figures 1-6**). Additional Catchup mice were used for the high spatial resolution imaging experiments (**Figure 7**). For the main dataset, the 9 infected sections (3 section replicates for 3 biological replicates-mice) are located in \"imzmls-Infected\" folder. The 9 mock-infected sections are located in \"imzmls-m...