Dataset Information

Flow cytometry has a significant impact on the cellular metabolome (Lipidomic LC-MS assay)

ABSTRACT: The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyse the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites related to the plasma membrane, activating a mechanosensory signalling cascade causing inflammation-like stress. The procedure also triggered alterations related to energy consumption and cell damage. The observed changes mostly derive from the physical impact on cells during their passage through the instrument. These findings provide evidence of FACS-induced biochemical changes, which should be taken into account in the design of robust metabolic assays of cells separated by flow cytometry.

The Lipidomic LC-MS assay is reported in the current study MTBLS631.
The General LC-MS assay is reported in MTBLS629.
The CE-MS assay is reported in MTBLS633.
The GC-MS assay is reported in MTBLS634.

Linked Studies: MTBLS629 MTBLS633 MTBLS634

INSTRUMENT(S): 6550 iFunnel Q-TOF LC/MS (Agilent)

SUBMITTER: Aleksandra Binek

PROVIDER: MTBLS631 | MetaboLights | 2018-11-06

REPOSITORIES: MetaboLights

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Publications

Flow Cytometry Has a Significant Impact on the Cellular Metabolome.

Binek Aleksandra A Rojo David D Godzien Joanna J Rupérez Francisco Javier FJ Nuñez Vanessa V Jorge Inmaculada I Ricote Mercedes M Vázquez Jesús J Barbas Coral C

Journal of proteome research 20181102 1

The characterization of specialized cell subpopulations in a heterogeneous tissue is essential for understanding organ function in health and disease. A popular method of cell isolation is fluorescence-activated cell sorting (FACS) based on probes that bind surface or intracellular markers. In this study, we analyze the impact of FACS on the cell metabolome of mouse peritoneal macrophages. Compared with directly pelleted macrophages, FACS-treated cells had an altered content of metabolites relat ...[more]

PMID: 30362351

Similar Datasets

Project description:Lipid Data Analyzer (LDA; http://genome.tugraz.at/lda2) provides automated and reliable annotation of lipid species and their molecular structures in high-throughput data from chromatography-coupled tandem mass spectrometry using embedded rule sets. Using various low- and high-resolution mass spectrometry instruments with several collision energies, the method's platform independence is proved. The software's reliability, flexibility, and ability to identify novel lipid molecular species may now render current state-of-the-art lipid libraries obsolete. LDA identified several novel lipid molecular species and novel regio-isomeric species; we consider a lipid species as “novel” if it is neither present in LIPID MAPS Structure Database, ChEBI, CyberLipid, HMDB, nor YMDB. Where evidence relies on fragments of very small intensity, high resolution HCD mode was used for final characterization. These data are presented here (CID mode data can be found in MTBLS397 - both biological experiments for Orbitrap Velos CID). Moreover, we present in this study data showing the LDA's ability to distinguish regio-isomeric standards under certain chromatographic conditions. Control experiment 1 assays for this study can be found in the MetaboLights study MTBLS394. Control experiment 2 assays for this study can be found in the MetaboLights study MTBLS391.Control experiment 3 assays for this study can be found in the MetaboLights study MTBLS398.Murine liver lipidome experiment assays for this study can be found in the MetaboLights study MTBLS396.LipidBlast benchmarking assays for this study can be found in the MetaboLights study MTBLS397. Linked Studies:<a href='https://www.ebi.ac.uk/metabolights/MTBLS391' target='_blank'>MTBLS391</a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS394' target='_blank'>MTBLS394</a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS396' target='_blank'>MTBLS396</a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS397' target='_blank'>MTBLS397</a> <a href='https://www.ebi.ac.uk/metabolights/MTBLS398' target='_blank'>MTBLS398</a>

			Action	DRS
	MTBLS631	Other
	FILES	Other
	a_MTBLS631_NEG_mass_spectrometry.txt	Txt
	a_MTBLS631_POS_mass_spectrometry.txt	Txt
	i_Investigation.txt	Txt