Proteomics

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Single-cell Proteomics of Developing Cells in Live Vertebrate Embryos by Microprobe High-Resolution Mass Spectrometry

ABSTRACT: In this work, we developed a microprobe capillary electrophoresis high-resolution mass spectrometry approach to identify proteins in single identified cells in live embryos of the South African clawed frog (Xenopus laevis) and also zebrafish. We used a pulled borosilicate capillary to aspirate a calibrated portion of the cell of interest. Proteins in the collected aspirate were detected and quantified via a bottom-up proteomic workflow that was downscaled to the single cells. The microprobe mass spectrometry platform is scalable to smaller cells in live, morphologically complex, developing embryos. As an example, we used this approach to analyze proteomic changes in cells that form a clone in the early developing Xenopus laevis embryo.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Danio Rerio (zebrafish) (brachydanio Rerio) Xenopus Laevis (african Clawed Frog)

TISSUE(S): Embryo, Stem Cell

SUBMITTER: Peter Nemes  

LAB HEAD: Peter Nemes

PROVIDER: PXD006905 | Pride | 2020-05-26

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
2016-07-27_CL02_nESI_D11_u-AmmBic_1-1.RAW Raw
2016-07-27_CL03_nESI_D11_u-AmmBic_1-2.RAW Raw
2016-07-27_CL04_nESI_D11_u-AmmBic_1-3.RAW Raw
2016-08-03_CL02_nESI_D11_usample-AmmBic_5-1.RAW Raw
2016-08-03_CL03_nESI_D11_usample-AmmBic_5-2.RAW Raw
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Publications

Microsampling Capillary Electrophoresis Mass Spectrometry Enables Single-Cell Proteomics in Complex Tissues: Developing Cell Clones in Live Xenopus laevis and Zebrafish Embryos.

Lombard-Banek Camille C   Moody Sally A SA   Manzini M Chiara MC   Nemes Peter P  

Analytical chemistry 20190318 7

Label-free single-cell proteomics by mass spectrometry (MS) is currently incompatible with complex tissues without requiring cell culturing, single-cell dissection, or tissue dissociation. We here report the first example of label-free single-cell MS-based proteomics directly in single cells in live vertebrate embryos. Our approach integrates optically guided in situ subcellular capillary microsampling, one-pot extraction-digestion of the collected proteins, peptide separation by capillary elect  ...[more]

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