Cited 15 time in webofscience Cited 15 time in scopus

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

Title
In Situ metabolic analysis of single plant cells by capillary microsampling and electrospray ionization mass spectrometry with ion mobility separation
Authors
Zhang, LinwenForeman, Daniel P.Grant, Paaqua A.Shrestha, BindeshMoody, Sally A.Villiers, FlorentKwak, June M.Vertes, Akos
DGIST Authors
Kwak, June M.
Issue Date
2014-10-21
Citation
Analyst, 139(20), 5079-5085
Type
Article
Article Type
Article
Keywords
ArabidopsisChemistryCytologyDevicesElectrospray Mass SpectrometryIsomerismMetabolismMetabolomicsPlant CellPlant CellsPlant LeafPlant LeavesProceduresSingle-Cell AnalysisSpectrometry, Mass, Electrospray Ionization
ISSN
0003-2654
Abstract
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.
URI
http://hdl.handle.net/20.500.11750/3018
DOI
10.1039/c4an01018c
Publisher
Royal Society of Chemistry
Related Researcher
  • Author Kwak, June Myoung KWAK_LAB(Lab of Cell Signailng)
  • Research Interests Calcium Signaling; ABA;ROS Signaling; drought-tolerant plants; Development of Abscission; Culluar Features; Signaling network; Environmental Regulation
Files:
There are no files associated with this item.
Collection:
New BiologyETC1. Journal Articles
New BiologyKWAK_LAB(Lab of Cell Signailng)1. Journal Articles


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