title: MALDI-guided SIMS: multiscale imaging of metabolites in bacterial biofilms.
abstract: Mass spectrometry imaging (MSI) is a versatile tool for visualizing molecular distributions in complex biological specimens, but locating microscopic chemical features of interest can be challenging in samples that lack a well-defined anatomy. To address this issue, we developed a correlated imaging approach that begins with performing matrix-assisted laser desorption/ionization (MALDI) MSI to obtain low-resolution molecular maps of a sample. The resulting maps are then used to direct subsequent microscopic secondary ion mass spectrometry (SIMS) imaging and tandem mass spectrometry (MS/MS) experiments to examine selected chemical regions of interest. By employing MALDI undersampling, the sample surface is left mostly unperturbed and available for the SIMS analysis, while also generating an ablation array that can be used for navigation in SIMS. We validated this MALDI-guided SIMS approach using cultured biofilms of the opportunistic pathogen Pseudomonas aeruginosa; bioactive secondary metabolites, including rhamnolipids and quinolones, were detected and visualized on both macro- and microscopic size scales. MSI mass assignments were confirmed with in situ MALDI MS/MS and capillary electrophoresis-electrospray ionization MS/MS analysis of biofilm extracts. Two strains of P. aeruginosa were compared, wild type and a quorum sensing mutant, and differences in metabolite abundance and distribution were observed.
authors.affiliation: Department of Chemistry and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Lanni
authors.forename: Rachel N
authors.initials: RN
authors.lastname: Masyuko
authors.forename: Callan M
authors.initials: CM
authors.lastname: Driscoll
authors.forename: Jordan T
authors.initials: JT
authors.lastname: Aerts
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Shrout
authors.forename: Paul W
authors.initials: PW
authors.lastname: Bohn
authors.forename: Jonathan V
authors.initials: JV
authors.lastname: Sweedler
mesh_terms: D018441:Biofilms / Q000254:growth & development*; D019075:Electrophoresis, Capillary; D006017:Glycolipids / Q000032:analysis; D055442:Metabolome; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D015363:Quinolones / Q000032:analysis; D053038:Quorum Sensing / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018629:Spectrometry, Mass, Secondary Ion
chemical_list: D006017:Glycolipids; D015363:Quinolones; C418382:rhamnolipid