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

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...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2019-04, Vol.91 (7), p.4797-4805
Main Authors: Lombard-Banek, Camille, Moody, Sally A, Manzini, M. Chiara, Nemes, Peter
Format: Article
Language:English
Subjects:
Amphibians
Animals
Capillary electrophoresis
Chemistry
Clone Cells - chemistry
Clone Cells - cytology
Electrokinetics
Electrophoresis
Electrophoresis, Capillary
Embryo, Nonmammalian - chemistry
Embryo, Nonmammalian - cytology
Embryos
Ionization
Mass Spectrometry
Mass spectroscopy
Proteins
Proteomics
Scientific imaging
Single-Cell Analysis
Spectroscopy
Technology
Tissues
Vertebrates
Xenopus laevis
Xenopus Proteins - analysis
Zebrafish
Zebrafish Proteins - analysis
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Summary: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 electrophoresis, ionization by an ultrasensitive electrokinetically pumped nanoelectrospray, and detection by high-resolution MS (Orbitrap). With a 700 zmol (420 000 copies) lower limit of detection, this trace-sensitive technology confidently identified and quantified ∼750–800 protein groups (
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.9b00345