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Download Hi-Res ImageDownload to MS-PowerPointCite This:Anal. Chem. 2019, 91, 9, 6118-6126
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    Rapid, Untargeted Chemical Profiling of Single Cells in Their Native Environment
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    • John F. Cahill*
      John F. Cahill
      Mass Spectrometry and Laser Spectroscopy Group, Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831-6131, United States
      *E-mail: [email protected]. Phone: 865-574-4878.
      More by John F. Cahill
      Orcidhttp://orcid.org/0000-0002-9866-4010
    • Julian Riba
      Julian Riba
      Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg, Georges-Köhler-Allee 103, 79110 Freiburg, Germany
      Cytena GmbH, Neuer Messplatz 3, 79108 Freiburg, Germany
      More by Julian Riba
    • Vilmos Kertesz
      Vilmos Kertesz
      Mass Spectrometry and Laser Spectroscopy Group, Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831-6131, United States
      More by Vilmos Kertesz
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    Analytical Chemistry

    Cite this: Anal. Chem. 2019, 91, 9, 6118–6126
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    https://doi.org/10.1021/acs.analchem.9b00680
    Published April 8, 2019
    Copyright © 2019 American Chemical Society
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    Abstract

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    We report a method that enables untargeted, high throughput, and quantitative mass spectrometric analysis of single cells from cell suspension without needing additional sample preparation procedures (e.g., molecular tagging) through the combination of single-cell printer technology and liquid vortex capture–mass spectrometry (SCP-LVC-MS). The operating principle behind the SCP-LVC-MS technology is single cell isolation via small droplet piezoelectric ejection followed by capture of the droplet into an LVC-MS sampling probe. Once exposed to an appropriate solvent, the cell is lysed, extracted, and analyzed by MS. The SCP-LVC-MS approach was validated by measuring the lipid composition of microalgae, Chlamydomonas reinhardtii (ChRe) and Euglena gracilis (EuGr), and HeLa cells in their native growth media. Numerous diacylglyceryltrimethylhomo-Ser (DGTS), phosphatidylcholine (PC), monogalactosyldiacylglycerol (MGDG), and digalactosyldiacylglycerol (DGDG) lipids were observed in single cells. Continuous solvent flow ensures that cells are analyzed rapidly, and no signal carryover between cells is observed. ChRe and EuGr microalgae mixed together in the same solution were differentiated cell-by-cell in real-time based on differences between levels of diacylglyceryltrimethylhomo-Ser (DGTS) and phosphatidylcholine (PC) lipids measured in each cell. Several DGTS lipids present in ChRe were quantified with single-cell resolution by normalizing to a DGTS(32:0) internal standard added to the LVC probe solvent during analysis. Quantitative peak areas were validated by comparing to bulk lipid extracts. Lastly, peak area distributions comprised of hundreds of cells were compared for ChRe after 5 days of nitrogen-limited and normal growth conditions, which show clear differences and the ability to resolve cellular population differences with single-cell resolution.

    Copyright © 2019 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.analchem.9b00680.

    • SCP-LVC-MS methods, Bligth and Dyer extraction protocol, cell ejection time lapse, caffeine sampling, high throughput single cell sampling, lysis comparison of cell wall and wall-less mutant (PDF)

    • Video of SCP-LVC-MS sampling (AVI)

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    Cited By

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    This article is cited by 46 publications.

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    Analytical Chemistry

    Cite this: Anal. Chem. 2019, 91, 9, 6118–6126
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acs.analchem.9b00680
    Published April 8, 2019
    Copyright © 2019 American Chemical Society
    Request reuse permissions

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