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The quantification of small molecules in single cells raises new potentials for better understanding the basic processes that underlie embryonic development. To enable single-cell investigations directly in live embryos, new analytical approaches are needed, particularly those that are sensitive, selective, quantitative, robust, and scalable to different cell sizes. Here, we present a protocol that enables the in situ analysis of metabolism in single cells in freely developing embryos of the South African clawed frog (Xenopus laevis), a powerful model in cell and developmental biology. This approach uses a capillary microprobe to aspirate a defined portion from single identified cells in the embryo, leaving neighboring cells intact for subsequent analysis. The collected cell content is analyzed by a microscale capillary electrophoresis electrospray ionization (CE-ESI) interface coupled to a high-resolution tandem mass spectrometer. This approach is scalable to various cell sizes and compatible with the complex three-dimensional structure of the developing embryo. As an example, we demonstrate that microprobe single-cell CE-ESI-MS enables the elucidation of metabolic cell heterogeneity that unfolds as a progenitor cell gives rise to descendants during development of the embryo. Besides cell and developmental biology, the single-cell analysis protocols described here are amenable to other cell sizes, cell types, or animal models.
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???displayArticle.pmcLink???PMC5755674 ???displayArticle.link???J Vis Exp ???displayArticle.grants???[+]
Aerts,
Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization.
2014, Pubmed
Aerts,
Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization.
2014,
Pubmed
Bodenmiller,
Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators.
2012,
Pubmed
Bowes,
Xenbase: a Xenopus biology and genomics resource.
2008,
Pubmed
,
Xenbase
Comi,
Categorizing Cells on the Basis of their Chemical Profiles: Progress in Single-Cell Mass Spectrometry.
2017,
Pubmed
Dale,
Fate map for the 32-cell stage of Xenopus laevis.
1987,
Pubmed
,
Xenbase
Gholipour,
In Situ Pressure Probe Sampling and UV-MALDI MS for Profiling Metabolites in Living Single Cells.
2012,
Pubmed
Grant,
Blastomere explants to test for cell fate commitment during embryonic development.
2013,
Pubmed
,
Xenbase
Guillaume-Gentil,
Single-Cell Mass Spectrometry of Metabolites Extracted from Live Cells by Fluidic Force Microscopy.
2017,
Pubmed
Hu,
Synchronized Polarization Induced Electrospray: Comprehensively Profiling Biomolecules in Single Cells by Combining both Positive-Ion and Negative-Ion Mass Spectra.
2016,
Pubmed
Huhn,
Relevance and use of capillary coatings in capillary electrophoresis-mass spectrometry.
2010,
Pubmed
James-Zorn,
Xenbase: expansion and updates of the Xenopus model organism database.
2013,
Pubmed
,
Xenbase
Karpinka,
Xenbase, the Xenopus model organism database; new virtualized system, data types and genomes.
2015,
Pubmed
,
Xenbase
Klein,
The first cleavage furrow demarcates the dorsal-ventral axis in Xenopus embryos.
1987,
Pubmed
,
Xenbase
Klepárník,
Recent advances in the development of single cell analysis--a review.
2013,
Pubmed
Lanni,
Mass spectrometry imaging and profiling of single cells.
2012,
Pubmed
Lapainis,
Capillary electrophoresis with electrospray ionization mass spectrometric detection for single-cell metabolomics.
2009,
Pubmed
Liu,
Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry.
2014,
Pubmed
Lombard-Banek,
New-generation mass spectrometry expands the toolbox of cell and developmental biology.
2017,
Pubmed
,
Xenbase
Mizuno,
Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification.
2008,
Pubmed
Moini,
Ultrafast capillary electrophoresis/mass spectrometry with adjustable porous tip for a rapid analysis of protein digest in about a minute.
2014,
Pubmed
Moody,
Fates of the blastomeres of the 16-cell stage Xenopus embryo.
1987,
Pubmed
,
Xenbase
Moody,
Fates of the blastomeres of the 32-cell-stage Xenopus embryo.
1987,
Pubmed
,
Xenbase
Moody,
Cell lineage analysis in Xenopus embryos.
2000,
Pubmed
,
Xenbase
Nakashima,
Single-Cell Metabolite Profiling of Stalk and Glandular Cells of Intact Trichomes with Internal Electrode Capillary Pressure Probe Electrospray Ionization Mass Spectrometry.
2016,
Pubmed
Nemes,
Qualitative and quantitative metabolomic investigation of single neurons by capillary electrophoresis electrospray ionization mass spectrometry.
2013,
Pubmed
,
Xenbase
Nemes,
Metabolic differentiation of neuronal phenotypes by single-cell capillary electrophoresis-electrospray ionization-mass spectrometry.
2011,
Pubmed
Nemes,
Spraying mode effect on droplet formation and ion chemistry in electrosprays.
2007,
Pubmed
Onjiko,
Metabolic Comparison of Dorsal versus Ventral Cells Directly in the Live 8-cell Frog Embryo by Microprobe Single-cell CE-ESI-MS.
2017,
Pubmed
,
Xenbase
Onjiko,
Single-cell mass spectrometry reveals small molecules that affect cell fates in the 16-cell embryo.
2015,
Pubmed
,
Xenbase
Onjiko,
Single-cell mass spectrometry with multi-solvent extraction identifies metabolic differences between left and right blastomeres in the 8-cell frog (Xenopus) embryo.
2016,
Pubmed
,
Xenbase
Onjiko,
In Situ Microprobe Single-Cell Capillary Electrophoresis Mass Spectrometry: Metabolic Reorganization in Single Differentiating Cells in the Live Vertebrate (Xenopus laevis) Embryo.
2017,
Pubmed
,
Xenbase
Pan,
The single-probe: a miniaturized multifunctional device for single cell mass spectrometry analysis.
2014,
Pubmed
Rollman,
Ultrafast capillary electrophoresis/mass spectrometry of controlled substances with optical isomer separation in about a minute.
2016,
Pubmed
Rubakhin,
Profiling metabolites and peptides in single cells.
2011,
Pubmed
,
Xenbase
Rubakhin,
Progress toward single cell metabolomics.
2013,
Pubmed
Saha-Shah,
Segmented flow sampling with push-pull theta pipettes.
2016,
Pubmed
Sellick,
Metabolite extraction from suspension-cultured mammalian cells for global metabolite profiling.
2011,
Pubmed
Svatoš,
Single-cell metabolomics comes of age: new developments in mass spectrometry profiling and imaging.
2011,
Pubmed
Tang,
Development and applications of single-cell transcriptome analysis.
2011,
Pubmed
Tran,
Characterization of the imprinting signature of mouse embryo fibroblasts by RNA deep sequencing.
2014,
Pubmed
Veselovska,
Deep sequencing and de novo assembly of the mouse oocyte transcriptome define the contribution of transcription to the DNA methylation landscape.
2015,
Pubmed
Wang,
Single cell analysis: the new frontier in 'omics'.
2010,
Pubmed
Wishart,
HMDB 3.0--The Human Metabolome Database in 2013.
2013,
Pubmed
Zenobi,
Single-cell metabolomics: analytical and biological perspectives.
2013,
Pubmed
Zhang,
In situ metabolic analysis of single plant cells by capillary microsampling and electrospray ionization mass spectrometry with ion mobility separation.
2014,
Pubmed
Zhang,
Energy Charge, Redox State, and Metabolite Turnover in Single Human Hepatocytes Revealed by Capillary Microsampling Mass Spectrometry.
2015,
Pubmed
Zhu,
Liquid chromatography quadrupole time-of-flight mass spectrometry characterization of metabolites guided by the METLIN database.
2013,
Pubmed
