Abstract
Live single-cell mass spectrometry (LSC-MS) allows for the detection of hundreds to thousands of metabolite peaks acquired from a single plant cell within a few minutes. Plant cells are first observed under a stereomicroscope, a cell of interest is chosen, and then sampled using a metal-coated glass microcapillary for subsequent analysis. A few microliters of ionization solvent is then added to the rear end of the capillary followed by the introduction of the capillary’s content directly into the mass spectrometer. High voltage is applied between the capillary and the mass spectrometer inlet to induce nanospray ionization. Metabolite structural confirmation is performed using tandem mass spectrometry analysis (MS/MS) and fragments are matched with MS/MS databases to predict metabolic pathways. This method enables swift and direct molecular detection and identification of specific metabolites from a single plant cell along with their localization within the cell, which will allow for comprehensive understanding of plant metabolomics on a single cell level.
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Acknowledgments
We thank Dr. Patrick J. Rudewicz, Novartis Institutes for BioMedical Research, for his support with writing this chapter. This work was conducted under the RIKEN Pioneering Projects of “Single Cell Science,” “Integrated Lipidology,” and “Biology of Symbiosis” and fund for the Development of Advanced Measurement and Analysis System (SENTAN), by the Japan Agency for Medical Research and Development (AMED).
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Masuda, K., Abouleila, Y., Ali, A., Yanagida, T., Masujima, T. (2018). Live Single-Cell Mass Spectrometry (LSC-MS) for Plant Metabolomics. In: António, C. (eds) Plant Metabolomics. Methods in Molecular Biology, vol 1778. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7819-9_19
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DOI: https://doi.org/10.1007/978-1-4939-7819-9_19
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