Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections
Abstract Glycans play an important role in biology with multiple cellular functions ranging from cell signaling, mobility and growth to protein folding and localization. The N-glycosylation state within a tissue has been found to vary greatly between healthy and diseased patients and has proven to h...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-29560-6 |
| _version_ | 1827984230000361472 |
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| author | Andrej Grgic Kasper K. Krestensen Ron M. A. Heeren |
| author_facet | Andrej Grgic Kasper K. Krestensen Ron M. A. Heeren |
| author_sort | Andrej Grgic |
| collection | DOAJ |
| description | Abstract Glycans play an important role in biology with multiple cellular functions ranging from cell signaling, mobility and growth to protein folding and localization. The N-glycosylation state within a tissue has been found to vary greatly between healthy and diseased patients and has proven to have an important clinical diagnostic value. Matrix assisted laser-desorption ionization (MALDI) mass spectrometry imaging (MSI) allows for untargeted analysis of biomolecules, including N-glycans, on a tissue section and provides a spatial context of the analyte. Until now, N-glycans have been predominantly analyzed using MALDI MSI on formalin-fixed paraffin embedded (FFPE) tissue sections, however this greatly reduces the clinical applicability, as the FFPE embedding process alters the biological environment of the tissue. Here we developed a protocol that allows for MALDI MSI of N-glycans from fresh frozen tissue that matches the current standard of FFPE analysis. By optimizing several steps in the sample preparation, we see orders of magnitude increase in signal intensity. Furthermore, this method limits delocalization of released N-glycans, thus improving the effective spatial resolution of the label-free molecular images. This protocol provides a novel perspective towards clinical application of MALDI MSI and capitalizes on the diagnostic value of N-glycan analysis. |
| first_indexed | 2024-04-09T22:57:39Z |
| format | Article |
| id | doaj.art-be3a420f7d6047a48ee0e21f161be62e |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-09T22:57:39Z |
| publishDate | 2023-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-be3a420f7d6047a48ee0e21f161be62e2023-03-22T11:10:29ZengNature PortfolioScientific Reports2045-23222023-02-011311710.1038/s41598-023-29560-6Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sectionsAndrej Grgic0Kasper K. Krestensen1Ron M. A. Heeren2The Maastricht MultiModal Molecular Imaging (M4I) Institute, Division of Imaging Mass Spectrometry (IMS), Maastricht UniversityThe Maastricht MultiModal Molecular Imaging (M4I) Institute, Division of Imaging Mass Spectrometry (IMS), Maastricht UniversityThe Maastricht MultiModal Molecular Imaging (M4I) Institute, Division of Imaging Mass Spectrometry (IMS), Maastricht UniversityAbstract Glycans play an important role in biology with multiple cellular functions ranging from cell signaling, mobility and growth to protein folding and localization. The N-glycosylation state within a tissue has been found to vary greatly between healthy and diseased patients and has proven to have an important clinical diagnostic value. Matrix assisted laser-desorption ionization (MALDI) mass spectrometry imaging (MSI) allows for untargeted analysis of biomolecules, including N-glycans, on a tissue section and provides a spatial context of the analyte. Until now, N-glycans have been predominantly analyzed using MALDI MSI on formalin-fixed paraffin embedded (FFPE) tissue sections, however this greatly reduces the clinical applicability, as the FFPE embedding process alters the biological environment of the tissue. Here we developed a protocol that allows for MALDI MSI of N-glycans from fresh frozen tissue that matches the current standard of FFPE analysis. By optimizing several steps in the sample preparation, we see orders of magnitude increase in signal intensity. Furthermore, this method limits delocalization of released N-glycans, thus improving the effective spatial resolution of the label-free molecular images. This protocol provides a novel perspective towards clinical application of MALDI MSI and capitalizes on the diagnostic value of N-glycan analysis.https://doi.org/10.1038/s41598-023-29560-6 |
| spellingShingle | Andrej Grgic Kasper K. Krestensen Ron M. A. Heeren Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections Scientific Reports |
| title | Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections |
| title_full | Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections |
| title_fullStr | Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections |
| title_full_unstemmed | Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections |
| title_short | Optimized protocol for MALDI MSI of N-glycans using an on-tissue digestion in fresh frozen tissue sections |
| title_sort | optimized protocol for maldi msi of n glycans using an on tissue digestion in fresh frozen tissue sections |
| url | https://doi.org/10.1038/s41598-023-29560-6 |
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