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Ion mobility (IM) spectrometry is a robust method that allows for the rapid separation and detection of a wide range of compounds. In this technique, ions in the drift cell obtain an average drift velocity from an electric field based upon their collision cross section (W) or shape, which allows for the separation of molecules depending upon their three-dimensional structure. Combining matrix-assisted laser desorption/ionization (MALDI) mass spectrometry with IM results in the fast sorting of biomolecules in complex mixtures along trend lines. In this 2D analysis of biological families, lipids, peptides, and nucleotides are separated from each other by differences in their ion mobility drift times in a timescale of hundreds of microseconds. Molecular ions of similar chemical type fall along trend lines when plotted in two-dimensional plots of ion mobility drift time as a function of m/z. We have analyzed a wide variety of samples using MALDI-IM MS. This talk will highlight the use of MALDI-IM MS for the analysis of lipids and it advantages in mass spectrometry imaging.