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Issue 4, 2011
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The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

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Abstract

Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been used to create spatial distribution maps from lipids, peptides, and proteins in a variety of biological tissues. MALDI-MSI often involves trade-offs between the extent of analyte extraction and desired spatial resolution, compromises that can adversely affect detectability. For example, increasing the extraction time can lead to unwanted analyte spatial redistribution. With the stretched sample method (SSM), the extraction period can be extended, resulting in reduced analyte redistribution while suppressing detection of cationic salt adducts. The SSM involves thaw-mounting a thin tissue section onto a substrate of small glass beads embedded in Parafilm M and then stretching the membrane to fragment the tissue into thousands of bead-sized pieces. Here, we applied the SSM method to MALDI-MSI using rat spinal cord as a model. We used surface-modified beads coated with trypsin or chymotrypsin in order to facilitate controlled digestion and detection of proteins. The enzymatic reactions were maintained by repeatedly condensing water on the stretched sample surface. As a result, new peptides formed by tryptic or chymotryptic protein digestion were detected and identified using a combination of MALDI-MSI and offline liquid chromatography tandem mass spectrometric analysis. Localization of these peptides indicated the distribution of their proteins of origin, including myelin basic protein, actin beta, and tubulin alpha chain. Additionally, we used uncoated beads to create distribution maps of many endogenous lipids and small peptides. The extension of the SSM using modified beads resulted in the creation of mosaic bead surfaces where adjacent beads were coated with different enzymes or other reactive chemicals, permitting investigation of the distributions of a wider range of analytes in biological samples within a single experiment.

Graphical abstract: The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

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Publication details

The article was received on 09 Nov 2010, accepted on 16 Jan 2011 and first published on 01 Feb 2011


Article type: Edge Article
DOI: 10.1039/C0SC00563K
Citation: Chem. Sci., 2011,2, 785-795
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    The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

    K. R. Tucker, L. A. Serebryannyy, T. A. Zimmerman, S. S. Rubakhin and J. V. Sweedler, Chem. Sci., 2011, 2, 785
    DOI: 10.1039/C0SC00563K

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