Abstract
Matrix deposition is a critical step in tissue imaging by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). It greatly affects the quality of MALDI imaging, especially for the analytes (such as lipids) that may easily dissolve in the solvent used for the matrix application. This chapter describes the use of an oscillating capillary nebulizer (OCN) to spray small droplets of matrix aerosol onto the sample surface for improved matrix homogeneity, reduced crystal size, and controlled solvent effects. This protocol allows visualization of many different lipid species and, of particular interest, sphingolipids in tissue slices of Tay-Sachs/Sandhoff disease by imaging MALDI-MS. The structures of these lipids were identified by analysis of tissue extracts using electrospray ionization in conjunction with tandem mass spectrometry (MS/MS and MS3). These results illustrate the usefulness of tissue imaging MALDI-MS with matrix deposition by OCN for the molecular analysis in normal physiology and pathology. In addition, the observation of numerous lipid subclasses with distinct localizations in the brain slices demonstrates that imaging MALDI-MS could be effectively used for “lipidomic” studies.
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Acknowledgments
The authors would like to thank Drs. Richard Browner and Facundo Fernandez for providing the OCN sprayer, Dr. Markus Stoeckli for sharing the modified MMIST software, and Lan Sun for SEM analysis. This work is supported by NIH GM069338 (Lipid MAPS) and seed funding from Georgia Institute of Technology for the Mass Spectrometry Bio-Imaging Center.
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Chen, Y. et al. (2010). Imaging MALDI Mass Spectrometry of Sphingolipids Using an Oscillating Capillary Nebulizer Matrix Application System. In: Rubakhin, S., Sweedler, J. (eds) Mass Spectrometry Imaging. Methods in Molecular Biology, vol 656. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-746-4_7
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DOI: https://doi.org/10.1007/978-1-60761-746-4_7
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