Further, TIMS MSI provides another layer of molecular identification by providing analyte specific collisional cross section data along with accurate mass. For our analysis, we made use of this feature for identification of lipid features that were significantly different in the OPLS DA and univariate analysis. The m/z/CCS data obtained for each statistically different lipid compound were further validated through offline analysis of mouse brain tissue extracts using liquid chromatography coupled to ESI TIMS-TOF MS/MS. The TIMS MSI data (m/z/CCS) were matched against a generated database consisting of both the m/z/CCS and MS/MS data obtained by the LC-ESI TIMS experiments. Together this approach generated a list of confidently identified lipid species that were statistically significant in between the different groups compared including plaques vs background, plaque-core and plaque-periphery as well as 18-and 23-old mice.