title: N-Glycan matrix-assisted laser desorption/ionization mass spectrometry imaging protocol for formalin-fixed paraffin-embedded tissues.
abstract: BACKGROUND
  Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) of the proteome of a tissue has been an established technique for the past decade. In the last few years, MALDI-MSI of the N-glycome has emerged as a novel MALDI-MSI technique. To assess the accuracy and clinical significance of the N-linked glycan spatial distribution, we have developed a method that utilises MALDI-MSI followed by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) in order to assign glycan structures to the differentiating MALDI-MSI glycan masses released from the tissue glycoproteins.
  
  
  RESULTS
  Our workflow presents a comprehensive list of instructions on how to (i) apply MALDI-MSI to spatially map the N-glycome across formalin-fixed paraffin-embedded (FFPE) clinical samples, (ii) structurally characterise N-glycans extracted from consecutive FFPE tissue sections by LC/MS/MS, and (iii) match relevant N-glycan masses from MALDI-MSI with confirmed N-glycan structures determined by LC/MS/MS.
  
  
  CONCLUSIONS
  Our protocol provides groups that are new to this technique with instructions how to establish N-glycan MALDI-MSI in their laboratory. Furthermore, the method assigns N-glycan structural detail to the masses obtained in the MALDI-MS image. Copyright © 2017 John Wiley & Sons, Ltd.
authors.affiliation: Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide, Australia, 5005.|Institute for Photonics and Advanced Sensing (IPAS), University of Adelaide, Adelaide, Australia, 5005.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Briggs
authors.affiliation: Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide, Australia, 5005.
authors.forename: Yin Ying
authors.initials: YY
authors.lastname: Ho
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Pulau Pinang, Malaysia.
authors.forename: Gurjeet
authors.initials: G
authors.lastname: Kaur
authors.affiliation: Department of Gynaecological Oncology, Royal Adelaide Hospital, Adelaide, South Australia, 5005, Australia.|Robinson Institute, University of Adelaide, Adelaide, Australia, 5005.
authors.forename: Martin K
authors.initials: MK
authors.lastname: Oehler
authors.affiliation: ARC Centre for Nanoscale BioPhotonics (CNBP), University of Adelaide, Adelaide, Australia, 5005.|Biomolecular Frontiers Research Centre, Macquarie University, Sydney, Australia, 2109.
authors.forename: Arun V
authors.initials: AV
authors.lastname: Everest-Dass
authors.affiliation: ARC Centre for Nanoscale BioPhotonics (CNBP), University of Adelaide, Adelaide, Australia, 5005.|Biomolecular Frontiers Research Centre, Macquarie University, Sydney, Australia, 2109.
authors.forename: Nicolle H
authors.identifier: http://orcid.org/0000-0002-7532-4021
authors.initials: NH
authors.lastname: Packer
authors.affiliation: Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide, Australia, 5005.|Institute for Photonics and Advanced Sensing (IPAS), University of Adelaide, Adelaide, Australia, 5005.
authors.forename: Peter
authors.identifier: http://orcid.org/0000-0001-8493-2208
authors.initials: P
authors.lastname: Hoffmann