227
© Springer Nature B.V. 2020
G. Sindona et al. (eds.), Toxic Chemical and Biological Agents, NATO Science
for Peace and Security Series A: Chemistry and Biology,
https://doi.org/10.1007/978-94-024-2041-8_17
Chapter 17
Tri-Modal MALDI-MS Imaging
on the Same Tissue Section: Deeper
Molecular Insights of Disease
Vanna Denti, Sonia Guarnerio, Andrew Smith, Isabella Piga, Clizia Chinello,
and Fulvio Magni
17.1 Introduction
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging
(MSI) is a unique technology that enables the in-situ detection of a broad range of
biomolecules, including lipids, N-glycans, and proteins. In particular, combining
multilevel molecular data can potentially improve the detection of novel disease
signatures and lead to improved stratification of patients. Here, we present a
workflow that highlights the feasibility of detecting lipids, N-glycans, and proteins
from the same formalin-fixed paraffin embedded (FFPE) tissue section.
17.2 Methods
For each section of FFPE clinical specimens, MALDI-MSI was performed in three
modalities using a Bruker rapiflex™ MALDI tissuetyper™. Initially, deparaffinization, tissue rehydration and antigen retrieval were performed. Thus, lipids were
analysed within the m/z 400 to 1200 range. Subsequently, N-glycans, that were
cleaved following digestion with PNGase-F, were analysed within the m/z 1000 to
3000 range. Finally, tryptic peptides obtained following enzymatic digestion were
analysed within the m/z 700 to 3000 range. All image acquisitions were performed
using a laser beam scan of 44 μm and a raster setting of 50 μm in both x and y
coordinates.
V. Denti (*) · S. Guarnerio · A. Smith · I. Piga · C. Chinello · F. Magni
Clinical Proteomics and Metabolomics Unit, Department of Medicine and Surgery,
University of Milano-Bicocca, Milano, Italy
e-mail: v.denti@campus.unimib.it
© Springer Nature B.V. 2020
G. Sindona et al. (eds.), Toxic Chemical and Biological Agents, NATO Science
for Peace and Security Series A: Chemistry and Biology,
https://doi.org/10.1007/978-94-024-2041-8_17
Chapter 17
Tri-Modal MALDI-MS Imaging
on the Same Tissue Section: Deeper
Molecular Insights of Disease
Vanna Denti, Sonia Guarnerio, Andrew Smith, Isabella Piga, Clizia Chinello,
and Fulvio Magni
17.1 Introduction
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging
(MSI) is a unique technology that enables the in-situ detection of a broad range of
biomolecules, including lipids, N-glycans, and proteins. In particular, combining
multilevel molecular data can potentially improve the detection of novel disease
signatures and lead to improved stratification of patients. Here, we present a
workflow that highlights the feasibility of detecting lipids, N-glycans, and proteins
from the same formalin-fixed paraffin embedded (FFPE) tissue section.
17.2 Methods
For each section of FFPE clinical specimens, MALDI-MSI was performed in three
modalities using a Bruker rapiflex™ MALDI tissuetyper™. Initially, deparaffinization, tissue rehydration and antigen retrieval were performed. Thus, lipids were
analysed within the m/z 400 to 1200 range. Subsequently, N-glycans, that were
cleaved following digestion with PNGase-F, were analysed within the m/z 1000 to
3000 range. Finally, tryptic peptides obtained following enzymatic digestion were
analysed within the m/z 700 to 3000 range. All image acquisitions were performed
using a laser beam scan of 44 μm and a raster setting of 50 μm in both x and y
coordinates.
V. Denti (*) · S. Guarnerio · A. Smith · I. Piga · C. Chinello · F. Magni
Clinical Proteomics and Metabolomics Unit, Department of Medicine and Surgery,
University of Milano-Bicocca, Milano, Italy
e-mail: v.denti@campus.unimib.it