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MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms

[Image: see text] 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...

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Autores principales: Lanni, Eric J., Masyuko, Rachel N., Driscoll, Callan M., Aerts, Jordan T., Shrout, Joshua D., Bohn, Paul W., Sweedler, Jonathan V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4165220/
https://www.ncbi.nlm.nih.gov/pubmed/25133532
http://dx.doi.org/10.1021/ac5020222
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author Lanni, Eric J.
Masyuko, Rachel N.
Driscoll, Callan M.
Aerts, Jordan T.
Shrout, Joshua D.
Bohn, Paul W.
Sweedler, Jonathan V.
author_facet Lanni, Eric J.
Masyuko, Rachel N.
Driscoll, Callan M.
Aerts, Jordan T.
Shrout, Joshua D.
Bohn, Paul W.
Sweedler, Jonathan V.
author_sort Lanni, Eric J.
collection PubMed
description [Image: see text] 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.
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spelling pubmed-41652202014-09-17 MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms Lanni, Eric J. Masyuko, Rachel N. Driscoll, Callan M. Aerts, Jordan T. Shrout, Joshua D. Bohn, Paul W. Sweedler, Jonathan V. Anal Chem [Image: see text] 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. American Chemical Society 2014-08-18 2014-09-16 /pmc/articles/PMC4165220/ /pubmed/25133532 http://dx.doi.org/10.1021/ac5020222 Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html)
spellingShingle Lanni, Eric J.
Masyuko, Rachel N.
Driscoll, Callan M.
Aerts, Jordan T.
Shrout, Joshua D.
Bohn, Paul W.
Sweedler, Jonathan V.
MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title_full MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title_fullStr MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title_full_unstemmed MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title_short MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms
title_sort maldi-guided sims: multiscale imaging of metabolites in bacterial biofilms
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4165220/
https://www.ncbi.nlm.nih.gov/pubmed/25133532
http://dx.doi.org/10.1021/ac5020222
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