Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers
Abstract Single‐cell MALDI mass spectrometry imaging (MSI) of lipids and metabolites >200 Da has recently come to the forefront of biomedical research and chemical biology. However, cell‐targeting and metabolome‐preserving methods for analysis of low mass, hydrophilic metabolites (<200 Da) in...
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| Format: | Article |
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Wiley
2025-02-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202410506 |
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| author | James L. Cairns Johanna Huber Andrea Lewen Jessica Jung Stefan J. Maurer Tobias Bausbacher Stefan Schmidt Pavel A. Levkin Daniel Sevin Kerstin Göpfrich Philipp Koch Oliver Kann Carsten Hopf |
| author_facet | James L. Cairns Johanna Huber Andrea Lewen Jessica Jung Stefan J. Maurer Tobias Bausbacher Stefan Schmidt Pavel A. Levkin Daniel Sevin Kerstin Göpfrich Philipp Koch Oliver Kann Carsten Hopf |
| author_sort | James L. Cairns |
| collection | DOAJ |
| description | Abstract Single‐cell MALDI mass spectrometry imaging (MSI) of lipids and metabolites >200 Da has recently come to the forefront of biomedical research and chemical biology. However, cell‐targeting and metabolome‐preserving methods for analysis of low mass, hydrophilic metabolites (<200 Da) in large cell populations are lacking. Here, the PRISM‐MS (PRescan Imaging for Small Molecule – Mass Spectrometry) mass‐guided MSI workflow is presented, which enables space‐efficient single cell lipid and metabolite analysis. In conjunction with giant unilamellar vesicles (GUVs) as MSI ground truth for cell‐sized objects and Monte Carlo reference‐based consensus clustering for data‐dependent identification of cell subpopulations, PRISM‐MS enables MSI and on‐cell MS2‐based identification of low‐mass metabolites like amino acids or Krebs cycle intermediates involved in stimulus‐dependent cell activation. The utility of PRISM‐MS is demonstrated through the characterization of complex metabolome changes in lipopolysaccharide (LPS)‐stimulated microglial cells and human‐induced pluripotent stem cell‐derived microglia. Translation of single cell results to endogenous microglia in organotypic hippocampal slice cultures indicates that LPS‐activation involves changes of the itaconate‐to‐taurine ratio and alterations in neuron‐to‐glia glutamine‐glutamate shuttling. The data suggests that PRISM‐MS can serve as a standard method in single cell metabolomics, given its capability to characterize larger cell populations and low‐mass metabolites. |
| format | Article |
| id | doaj-art-12ffc1e80d0546b5b596b752361f6911 |
| institution | Kabale University |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-12ffc1e80d0546b5b596b752361f69112025-02-04T13:14:54ZengWileyAdvanced Science2198-38442025-02-01125n/an/a10.1002/advs.202410506Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation MarkersJames L. Cairns0Johanna Huber1Andrea Lewen2Jessica Jung3Stefan J. Maurer4Tobias Bausbacher5Stefan Schmidt6Pavel A. Levkin7Daniel Sevin8Kerstin Göpfrich9Philipp Koch10Oliver Kann11Carsten Hopf12Center for Mass Spectrometry and Optical Spectroscopy CeMOS Mannheim University of Applied Sciences 68163 Mannheim GermanyCenter for Mass Spectrometry and Optical Spectroscopy CeMOS Mannheim University of Applied Sciences 68163 Mannheim GermanyInstitute of Physiology and Pathophysiology Heidelberg University 69120 Heidelberg GermanyDept. Translational Brain Research Central Institute for Mental Health (CIMH) 68159 Mannheim GermanyBiophysical Engineering Group Center for Molecular Biology of Heidelberg University (ZMBH) 69120 Heidelberg GermanyCenter for Mass Spectrometry and Optical Spectroscopy CeMOS Mannheim University of Applied Sciences 68163 Mannheim GermanyCenter for Mass Spectrometry and Optical Spectroscopy CeMOS Mannheim University of Applied Sciences 68163 Mannheim GermanyInstitute of Biological and Chemical Systems – Functional Molecular Systems (IBCS‐FMS) Karlsruhe Institute of Technology 76344 Karlsruhe GermanyCellzome – A GSK company 69115 Heidelberg GermanyBiophysical Engineering Group Center for Molecular Biology of Heidelberg University (ZMBH) 69120 Heidelberg GermanyDept. Translational Brain Research Central Institute for Mental Health (CIMH) 68159 Mannheim GermanyInstitute of Physiology and Pathophysiology Heidelberg University 69120 Heidelberg GermanyCenter for Mass Spectrometry and Optical Spectroscopy CeMOS Mannheim University of Applied Sciences 68163 Mannheim GermanyAbstract Single‐cell MALDI mass spectrometry imaging (MSI) of lipids and metabolites >200 Da has recently come to the forefront of biomedical research and chemical biology. However, cell‐targeting and metabolome‐preserving methods for analysis of low mass, hydrophilic metabolites (<200 Da) in large cell populations are lacking. Here, the PRISM‐MS (PRescan Imaging for Small Molecule – Mass Spectrometry) mass‐guided MSI workflow is presented, which enables space‐efficient single cell lipid and metabolite analysis. In conjunction with giant unilamellar vesicles (GUVs) as MSI ground truth for cell‐sized objects and Monte Carlo reference‐based consensus clustering for data‐dependent identification of cell subpopulations, PRISM‐MS enables MSI and on‐cell MS2‐based identification of low‐mass metabolites like amino acids or Krebs cycle intermediates involved in stimulus‐dependent cell activation. The utility of PRISM‐MS is demonstrated through the characterization of complex metabolome changes in lipopolysaccharide (LPS)‐stimulated microglial cells and human‐induced pluripotent stem cell‐derived microglia. Translation of single cell results to endogenous microglia in organotypic hippocampal slice cultures indicates that LPS‐activation involves changes of the itaconate‐to‐taurine ratio and alterations in neuron‐to‐glia glutamine‐glutamate shuttling. The data suggests that PRISM‐MS can serve as a standard method in single cell metabolomics, given its capability to characterize larger cell populations and low‐mass metabolites.https://doi.org/10.1002/advs.202410506giant unilamellar vesicles (GUVs)human induced pluripotent stem cells (hiPSC)MALDI mass spectrometry imagingmicroglianeurodegenerationsingle cell |
| spellingShingle | James L. Cairns Johanna Huber Andrea Lewen Jessica Jung Stefan J. Maurer Tobias Bausbacher Stefan Schmidt Pavel A. Levkin Daniel Sevin Kerstin Göpfrich Philipp Koch Oliver Kann Carsten Hopf Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers Advanced Science giant unilamellar vesicles (GUVs) human induced pluripotent stem cells (hiPSC) MALDI mass spectrometry imaging microglia neurodegeneration single cell |
| title | Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers |
| title_full | Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers |
| title_fullStr | Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers |
| title_full_unstemmed | Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers |
| title_short | Mass‐Guided Single‐Cell MALDI Imaging of Low‐Mass Metabolites Reveals Cellular Activation Markers |
| title_sort | mass guided single cell maldi imaging of low mass metabolites reveals cellular activation markers |
| topic | giant unilamellar vesicles (GUVs) human induced pluripotent stem cells (hiPSC) MALDI mass spectrometry imaging microglia neurodegeneration single cell |
| url | https://doi.org/10.1002/advs.202410506 |
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