Development of Quantification Based MS Imaging

QUEST-MSI project aims to set up a novel analytical platform, for spatially-resolved, quantitative, and highly-sensitive analysis of proteins. A new workflow based on spatially resolved protein extraction followed by in-solution processing will allow high confidence identification of a large number of proteins, quantification of identified proteins, and the ability to reconstruct quantitative images of spatial abundance for the identified proteins.

QUEST-MSI: Proof-of-principle experiment of quantitative protein analysis of a half of a rat brain tissue section which demonstrates the complete workflow proposed in this project. Brain section was deposited on a parafilm and pixels were cut-out under microscope using a scalpel. Each pixel was processed separately for protein extraction, reduction/alkylation and trypsin digestion. Tryptic peptides were then separated and analyzed on a LC-ESI system; MS and MS2 data were stored. Recorded data processing allowed us for protein ID and for relative quantification using spectral counting under Scaffold. Image reconstruction was performed in the Microsoft Excel software (Franck et al., 2013).

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  • Franck J., Quanico J., Wisztorski M. , Day R., Salzet M. and Fournier I.
    Anal. Chem. 2013, 85: 8127−8134
    Quantification Based Mass Spectrometry Imaging of Proteins by Parafilm Assisted Microdissection
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  • Quanico J., Franck J., Dauly C;, Strupat K., Dupuy J., Day R., Salzet M., Fournier I., Wisztorski M.
    J. Proteomics 2013, 79: 200-218
    Development of Liquid Microjunction Extraction Strategy for Improving Protein Identification from Tissue Sections
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  • Wisztorski M., Fatou B., Franck J., desmons A., Farré, I., Leblanc E., Fournier I., Salzet M.
    Proteomic Clinical Applications 2013, 7(3-4): 234-40
    Microproteomics by Liquid Extraction Surface Analysis: Application to FFPE tissue to study the Fimbria region of tubo-ovarian cancer
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