Premium
N‐ Glycan matrix‐assisted laser desorption/ionization mass spectrometry imaging protocol for formalin‐fixed paraffin‐embedded tissues
Author(s) -
Briggs Matthew T.,
Ho Yin Ying,
Kaur Gurjeet,
Oehler Martin K.,
EverestDass Arun V.,
Packer Nicolle H.,
Hoffmann Peter
Publication year - 2017
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.7845
Subject(s) - glycan , chemistry , glycome , mass spectrometry , mass spectrometry imaging , chromatography , matrix assisted laser desorption/ionization , tandem mass spectrometry , maldi imaging , glycoprotein , desorption , biochemistry , organic chemistry , adsorption
Rationale 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. Methods and 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.