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Microprobe Capillary Electrophoresis Mass Spectrometry for Single-cell Metabolomics in Live Frog (Xenopus laevis) Embryos
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JoVE Journal Chemistry
Microprobe Capillary Electrophoresis Mass Spectrometry for Single-cell Metabolomics in Live Frog (Xenopus laevis) Embryos

Microprobe Capillary Electrophoresis Mass Spectrometry for Single-cell Metabolomics in Live Frog (Xenopus laevis) Embryos

DOI:
10.3791/56956-v

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12:16 min

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December 22, 2017

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, , ,
1Department of Chemistry,George Washington University, 2Department of Anatomy & Regenerative Biology,George Washington University, 3Department of Chemistry & Biochemistry,University of Maryland, College Park

Chapters

  • 00:05Title
  • 01:01Preparation of Sampling Instruments and Dishes
  • 02:01Microsampling Single Cells and Metabolite Extraction
  • 05:23Capillary Electrophoresis Electrospray Ionization Mass Spectrometry (CE-ESI-MS) Measurement
  • 09:01Results: Metabolic Differences Between Single Cells in the Early Developing Xenopus laevis Embryo
  • 10:43Conclusion

Summary

Automatic Translation

Automatic Translation

We describe steps that enable fast in situ sampling of a small portion of an individual cell with high precision and minimal invasion using capillary-based micro-sampling, to facilitate chemical characterization of a snapshot of metabolic activity in live embryos using a custom-built single cell capillary electrophoresis and mass spectrometry platform.

Tags

MicroprobeCapillary ElectrophoresisMass SpectrometrySingle-cell MetabolomicsLive FrogXenopus LaevisEmbryosIn Situ MicrosamplingCell BiologyDevelopmental BiologyMinimally Invasive SamplingAgarose GelPasteur PipetteBorosilicate CapillariesFlaming/Brown Type Capillary PullerDejellyingSteinberg’s SolutionGonadotropinIn Vitro Fertilization

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