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Single-cell mass spectrometry (MS) empowers metabolomic investigations by decreasing analytical dimensions to the size of individual cells and subcellular structures. We describe a protocol for investigating and quantifying metabolites in individual isolated neurons using single-cell capillary electrophoresis (CE) coupled to electrospray ionization (ESI) time-of-flight (TOF) MS. The protocol requires ∼2 h for sample preparation, neuron isolation and metabolite extraction, and 1 h for metabolic measurement. We used the approach to detect more than 300 distinct compounds in the mass range of typical metabolites in various individual neurons (25-500 μm in diameter) isolated from the sea slug (Aplysia californica) central and rat (Rattus norvegicus) peripheral nervous systems. We found that a subset of identified compounds was sufficient to reveal metabolic differences among freshly isolated neurons of different types and changes in the metabolite profiles of cultured neurons. The protocol can be applied to the characterization of the metabolome in a variety of smaller cells and/or subcellular domains.
Amantonico,
Analytical techniques for single-cell metabolomics: state of the art and trends.
2010, Pubmed
Amantonico,
Analytical techniques for single-cell metabolomics: state of the art and trends.
2010,
Pubmed
Baran,
MathDAMP: a package for differential analysis of metabolite profiles.
2006,
Pubmed
Barbula,
Interfacing capillary-based separations to mass spectrometry using desorption electrospray ionization.
2011,
Pubmed
Bodenmiller,
Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators.
2012,
Pubmed
Bonner,
Laser capture microdissection: molecular analysis of tissue.
1997,
Pubmed
Cecala,
Sampling techniques for single-cell electrophoresis.
2012,
Pubmed
Coello,
Atmospheric pressure femtosecond laser imaging mass spectrometry.
2010,
Pubmed
Cohen,
Chemical cytometry: fluorescence-based single-cell analysis.
2008,
Pubmed
Espina,
Laser-capture microdissection.
2006,
Pubmed
Eyer,
A microchamber array for single cell isolation and analysis of intracellular biomolecules.
2012,
Pubmed
Fischer,
Microscopy in 3D: a biologist's toolbox.
2011,
Pubmed
Geigl,
Single-cell isolation from cell suspensions and whole genome amplification from single cells to provide templates for CGH analysis.
2007,
Pubmed
Greving,
Nanostructure-initiator mass spectrometry metabolite analysis and imaging.
2011,
Pubmed
Harris,
Recent developments in ambient ionization techniques for analytical mass spectrometry.
2008,
Pubmed
Hell,
Far-field optical nanoscopy.
2007,
Pubmed
Kilkenny,
Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research.
2010,
Pubmed
Kurczy,
Mass spectrometry imaging of mating Tetrahymena show that changes in cell morphology regulate lipid domain formation.
2010,
Pubmed
Lapainis,
Contributions of capillary electrophoresis to neuroscience.
2008,
Pubmed
Lapainis,
Capillary electrophoresis with electrospray ionization mass spectrometric detection for single-cell metabolomics.
2009,
Pubmed
Lee,
Use of mass spectrometry for imaging metabolites in plants.
2012,
Pubmed
Li,
Peptides in the brain: mass spectrometry-based measurement approaches and challenges.
2008,
Pubmed
Lin,
Chemical analysis of single cells.
2011,
Pubmed
Mellors,
Integrated microfluidic device for automated single cell analysis using electrophoretic separation and electrospray ionization mass spectrometry.
2010,
Pubmed
Miao,
Rapid isolation of single malaria parasite-infected red blood cells by cell sorting.
2011,
Pubmed
Monroe,
Vitamin E imaging and localization in the neuronal membrane.
2005,
Pubmed
Nemes,
Metabolic differentiation of neuronal phenotypes by single-cell capillary electrophoresis-electrospray ionization-mass spectrometry.
2011,
Pubmed
Nemes,
Laser ablation electrospray ionization for atmospheric pressure, in vivo, and imaging mass spectrometry.
2007,
Pubmed
Nemes,
Spraying mode effect on droplet formation and ion chemistry in electrosprays.
2007,
Pubmed
Nemes,
Single-cell metabolomics: changes in the metabolome of freshly isolated and cultured neurons.
2012,
Pubmed
,
Xenbase
Neupert,
Targeted single-cell microchemical analysis: MS-based peptidomics of individual paraformaldehyde-fixed and immunolabeled neurons.
2012,
Pubmed
Neupert,
Mass spectrometric analysis of single identified neurons of an insect.
2005,
Pubmed
Northen,
Clathrate nanostructures for mass spectrometry.
2007,
Pubmed
Page,
Single-neuron analysis using CE combined with MALDI MS and radionuclide detection.
2002,
Pubmed
Patti,
Meta-analysis of untargeted metabolomic data from multiple profiling experiments.
2012,
Pubmed
Planchon,
Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination.
2011,
Pubmed
Pluskal,
MZmine 2: modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data.
2010,
Pubmed
Rubakhin,
Profiling signaling peptides in single mammalian cells using mass spectrometry.
2006,
Pubmed
Rubakhin,
Profiling metabolites and peptides in single cells.
2011,
Pubmed
,
Xenbase
Rubakhin,
Characterizing peptides in individual mammalian cells using mass spectrometry.
2007,
Pubmed
Rubakhin,
Progress toward single cell metabolomics.
2013,
Pubmed
Schoenherr,
CE-microreactor-CE-MS/MS for protein analysis.
2007,
Pubmed
Shrestha,
In situ cell-by-cell imaging and analysis of small cell populations by mass spectrometry.
2011,
Pubmed
Smith,
METLIN: a metabolite mass spectral database.
2005,
Pubmed
Southey,
NeuroPred: a tool to predict cleavage sites in neuropeptide precursors and provide the masses of the resulting peptides.
2006,
Pubmed
Stephens,
Light microscopy techniques for live cell imaging.
2003,
Pubmed
Stolee,
Laser-nanostructure interactions for ion production.
2012,
Pubmed
Tsuyama,
Mass spectrometry for cellular and tissue analyses in a very small region.
2011,
Pubmed
Urban,
Mass spectrometric method incorporating enzymatic amplification for attomole-level analysis of target metabolites in biological samples.
2010,
Pubmed
Wang,
Single cell analysis: the new frontier in 'omics'.
2010,
Pubmed
Wishart,
HMDB: a knowledgebase for the human metabolome.
2009,
Pubmed
Yan,
Nanowire-based single-cell endoscopy.
2011,
Pubmed
Yang,
Detection of characteristic distributions of phospholipid head groups and fatty acids on neurite surface by time-of-flight secondary ion mass spectrometry.
2010,
Pubmed
