In Situ Microprobe Single-Cell Capillary Electrophoresis Mass

Apr 22, 2017 - ... Electrophoresis Mass Spectrometry: Metabolic Reorganization in Single Differentiating Cells in the Live Vertebrate (Xenopus laevis)...
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In Situ Microprobe Single-Cell Capillary Electrophoresis Mass Spectrometry: Metabolic Reorganization in Single Differentiating Cells in the Live Vertebrate (Xenopus laevis) Embryo Rosemary M. Onjiko,† Erika P. Portero,† Sally A. Moody,‡ and Peter Nemes*,† †

Department of Chemistry and ‡Department of Anatomy and Regenerative Biology, The George Washington University, Washington, D.C., 20052, United States S Supporting Information *

ABSTRACT: Knowledge of single-cell metabolism would provide a powerful look into cell activity changes as cells differentiate to all the tissues of the vertebrate embryo. However, single-cell mass spectrometry technologies have not yet been made compatible with complex three-dimensional changes and rapidly decreasing cell sizes during early development of the embryo. Here, we bridge this technological gap by integrating capillary microsampling, microscale metabolite extraction, and capillary electrophoresis electrospray ionization mass spectrometry (CE-ESI-MS) to enable direct metabolic analysis of identified cells in the live frog embryo (Xenopus laevis). Microprobe CE-ESI-MS of