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In Situ metabolic analysis of single plant cells by capillary microsampling and electrospray ionization mass spectrometry with ion mobility separation

In Situ metabolic analysis of single plant cells by capillary microsampling and electrospray ionization mass spectrometry with ion mobility separation
Zhang, LinwenForeman, Daniel P.Grant, Paaqua A.Shrestha, BindeshMoody, Sally A.Villiers, FlorentKwak, June M.Vertes, Akos
DGIST Authors
Kwak, June M.
Issued Date
Article Type
ArabidopsisChemistryCytologyDevicesElectrospray Mass SpectrometryIsomerismMetabolismMetabolomicsPlant CellPlant CellsPlant LeafPlant LeavesProceduresSingle-Cell AnalysisSpectrometry, Mass, Electrospray Ionization
Advances in single cell analysis techniques have demonstrated cell-to-cell variability in both homogeneous and heterogeneous cell populations strengthening our understanding of multicellular organisms and individual cell behaviour. However, additional tools are needed for non-targeted metabolic analysis of live single cells in their native environment. Here, we combine capillary microsampling with electrospray ionization (ESI) mass spectrometry (MS) and ion mobility separation (IMS) for the analysis of various single A. thaliana epidermal cell types, including pavement and basal cells, and trichomes. To achieve microsampling of different cell types with distinct morphology, custom-tailored microcapillaries were used to extract the cell contents. To eliminate the isobaric interferences and enhance the ion coverage in single cell analysis, a rapid separation technique, IMS, was introduced that retained ions based on their collision cross sections. For each cell type, the extracted cell material was directly electrosprayed resulting in ∼200 peaks in ESI-MS and ∼400 different ions in ESI-IMS-MS, the latter representing a significantly enhanced coverage. Based on their accurate masses and tandem MS, 23 metabolites and lipids were tentatively identified. Our results indicated that profound metabolic differences existed between the trichome and the other two cell types but differences between pavement and basal cells were hard to discern. The spectra indicated that in all three A. thaliana cell types the phenylpropanoid metabolism pathway had high coverage. In addition, metabolites from the subpathway, sinapic acid ester biosynthesis, were more abundant in single pavement and basal cells, whereas compounds from the kaempferol glycoside biosynthesis pathway were present at significantly higher level in trichomes. Our results demonstrate that capillary microsampling coupled with ESI-IMS-MS captures metabolic differences between A. thaliana epidermal cell types, paving the way for the non-targeted analysis of single plant cells and subcellular compartments. This journal is © 2014 the Partner Organisations.
Royal Society of Chemistry
Related Researcher
  • 곽준명 Kwak, June Myoung 뉴바이올로지학과
  • Research Interests Calcium Signaling; ABA;ROS Signaling; plant development; Development of Abscission; Culluar Features; Signaling network; Environmental Regulation
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Department of New Biology Lab of Cell Signaling and Development 1. Journal Articles


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