title: On-Tissue Phospholipase C Digestion for Enhanced MALDI-MS Imaging of Neutral Glycosphingolipids.
abstract: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) can be used to simultaneously visualize the lateral distribution of different lipid classes in tissue sections, but the applicability of the method to real-life samples is often limited by ion suppression effects. In particular, the presence of abundant phosphatidylcholines (PCs) can reduce the ion yields for all other lipid species in positive ion mode measurements. Here, we used on-tissue treatment with buffer-free phospholipase C (PLC) to near-quantitatively degrade PCs in fresh-frozen tissue sections. The ion signal intensities of mono-, di-, and oligohexosylceramides were enhanced by up to 10-fold. In addition, visualization of Shiga toxin receptor globotriaosylceramide (Gb3Cer) in the kidneys of wild-type and α-galactosidase A-knockout (Fabry) mice was possible at about ten micrometer resolution. Importantly, the PLC treatment did not decrease the high lateral resolution of the MS imaging analysis.
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.|Interdisciplinary Center for Clinical Research (IZKF), University of Münster , 48149 Münster, Germany.
authors.forename: Simeon
authors.initials: S
authors.lastname: Vens-Cappell
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.|Interdisciplinary Center for Clinical Research (IZKF), University of Münster , 48149 Münster, Germany.
authors.forename: Ivan U
authors.initials: IU
authors.lastname: Kouzel
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.|Interdisciplinary Center for Clinical Research (IZKF), University of Münster , 48149 Münster, Germany.
authors.forename: Hans
authors.initials: H
authors.lastname: Kettling
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Soltwisch
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bauwens
authors.affiliation: Institute of Pathology Mannheim, University of Heidelberg , 68167 Mannheim, Germany.|Cellular and Molecular Pathology, German Cancer Research Center, DKFZ , 69120 Heidelberg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Porubsky
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.|Interdisciplinary Center for Clinical Research (IZKF), University of Münster , 48149 Münster, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Müthing
authors.affiliation: Institute for Hygiene, University of Münster , 48149 Münster, Germany.|Interdisciplinary Center for Clinical Research (IZKF), University of Münster , 48149 Münster, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Dreisewerd
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D007668:Kidney / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D020383:Neutral Glycosphingolipids / Q000032:analysis* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D010738:Type C Phospholipases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020383:Neutral Glycosphingolipids; D010738:Type C Phospholipases