Mass Spectrometry Imaging (MSI) of Fresh Bones using Infrared Matrix-Assisted Laser Desorption Electrospray Ionization (IR-MALDESI).

We report an effective strategy for direct analysis and two-dimensional (2D) matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) of mouse bones that underwent no chemical treatments prior to analysis. To unravel the chemistry in bones under near-physiological conditions, we cut a flash-frozen bone in half longitudinally, placed it in a mold facing flat side down, and poured Plaster of Paris on top of and around the bone. After Plaster of Paris had set, the bone with embedding material was removed from the mold, and placed on the IR-MALDESI imaging stage. Plaster of Paris acted as a fixture to keep every spot on the sample surface the same distance away from the laser focus. To demonstrate the feasibility of IR-MALDESI MSI for analyses of unmodified bones, we imaged bones derived from healthy and stroke-affected mice and generated ion heatmaps showing the spatial distribution of putatively annotated features.