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Dual Cationic-Anionic Profiling of Metabolites in a Single Embryonic Cell in a Live Xenopus laevis Embryo by Microprobe CE-ESI-MS

Abstract

In situ capillary microsampling with capillary electrophoresis (CE) electrospray ionization (ESI) mass spectrometry (MS) enabled characterization of cationic metabolites in single cells in complex tissues and organisms. For deeper coverage of the metabolome and metabolic networks, analytical approaches are needed that provide complementary detection for anionic metabolites, ideally using the same instrumentation. Described here is one such approach that enables sequential cationic and anionic (dual) analysis of the same identified cell in a live vertebrate embryo. A calibrated volume was microaspirated from the animal-dorsal cell in a live 8-cell embryo of Xenopus laevis, and cationic and anionic metabolites were one-pot microextracted from the aspirate, followed by CE-ESI-MS analysis of the same extract. A laboratory-built CE-ESI interface was reconfigured to enable dual cationic-anionic analysis with ~5–10 nM (50–100 amol) lower limit of detection and capability for quantification. To provide robust separation and efficient ion generation, the CE-ESI interface was enclosed in a nitrogen gas filled chamber, and the operational parameters were optimized for the cone-jet spraying regime in both the positive and negative ion mode. A total of ~250 cationic and ~200 anionic molecular features were detected from the cell between m/z 50–500, including 60 and 24 identified metabolites, respectively. With only 11 metabolites identified mutually, the duplexed approach yielded complementary information on metabolites produced in the cell, which in turn deepened network coverage for several metabolic pathways. With scalability to smaller cells and adaptability to other types of tissues other models, dual cationic-anionic detection with in situ CE-ESI-MS opens a door to better understand cell metabolism.

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Supplementary files

Publication details

The article was received on 17 Oct 2018, accepted on 05 Dec 2018 and first published on 05 Dec 2018


Article type: Paper
DOI: 10.1039/C8AN01999A
Citation: Analyst, 2018, Accepted Manuscript
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    Dual Cationic-Anionic Profiling of Metabolites in a Single Embryonic Cell in a Live Xenopus laevis Embryo by Microprobe CE-ESI-MS

    P. Nemes and E. P. Portero, Analyst, 2018, Accepted Manuscript , DOI: 10.1039/C8AN01999A

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