Von Gerichten, Johanna;
Saunders, Kyle DG;
Kontiza, Anastasia;
Newman, Carla F;
Mayson, George;
Beste, Dany JV;
Velliou, Eirini;
... Bailey, Melanie J; + view all
(2024)
Untargeted single-cell lipidomics using liquid chromatography and data-dependent acquisition after live cell selection.
Analytical Chemistry
, 96
(18)
pp. 6922-6929.
10.1021/acs.analchem.3c05677.
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Abstract
We report the development and validation of an untargeted single-cell lipidomics method based on microflow chromatography coupled to a data-dependent mass spectrometry method for fragmentation-based identification of lipids. Given the absence of single-cell lipid standards, we show how the methodology should be optimized and validated using a dilute cell extract. The methodology is applied to dilute pancreatic cancer and macrophage cell extracts and standards to demonstrate the sensitivity requirements for confident assignment of lipids and classification of the cell type at the single-cell level. The method is then coupled to a system that can provide automated sampling of live, single cells into capillaries under microscope observation. This workflow retains the spatial information and morphology of cells during sampling and highlights the heterogeneity in lipid profiles observed at the single-cell level. The workflow is applied to show changes in single-cell lipid profiles as a response to oxidative stress, coinciding with expanded lipid droplets. This demonstrates that the workflow is sufficiently sensitive to observing changes in lipid profiles in response to a biological stimulus. Understanding how lipids vary in single cells will inform future research into a multitude of biological processes as lipids play important roles in structural, biophysical, energy storage, and signaling functions.
| Type: | Article |
|---|---|
| Title: | Untargeted single-cell lipidomics using liquid chromatography and data-dependent acquisition after live cell selection |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acs.analchem.3c05677 |
| Publisher version: | https://doi.org/10.1021/acs.analchem.3c05677 |
| Language: | English |
| Additional information: | © The Author(s), 2024. This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. https://creativecommons.org/licenses/by/4.0/ |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Surgery and Interventional Sci UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Surgery and Interventional Sci > Department of Targeted Intervention |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10190390 |
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