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Subcellular-level resolution MALDI-MS imaging of maize leaf metabolites by MALDI-linear ion trap-Orbitrap mass spectrometer

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Abstract

A significant limiting factor in achieving high spatial resolution for matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) imaging is the size of the laser spot at the sample surface. Here, we present modifications to the beam-delivery optics of a commercial MALDI-linear ion trap-Orbitrap instrument, incorporating an external Nd:YAG laser, beam-shaping optics, and an aspheric focusing lens, to reduce the minimum laser spot size from ~50 μm for the commercial configuration down to ~9 μm for the modified configuration. This improved system was applied for MALDI-MS imaging of cross sections of juvenile maize leaves at 5-μm spatial resolution using an oversampling method. A variety of different metabolites including amino acids, glycerolipids, and defense-related compounds were imaged at a spatial resolution well below the size of a single cell. Such images provide unprecedented insights into the metabolism associated with the different tissue types of the maize leaf, which is known to asymmetrically distribute the reactions of C4 photosynthesis among the mesophyll and bundle sheath cell types. The metabolite ion images correlate with the optical images that reveal the structures of the different tissues, and previously known and newly revealed asymmetric metabolic features are observed.

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Acknowledgments

This work was supported by the US Department of Energy (DOE), Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences, and Biosciences. MDY-N acknowledges the support of the National Science Foundation under Award No. EEC-0813570 and Award No. IOS-1354799, which co-sponsored the development of the genetic stocks imaged in this study. The Ames Laboratory is operated by Iowa State University under DOE Contract DE-AC02-07CH11358.

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Authors and Affiliations

  1. Department of Chemistry, Iowa State University, Ames, IA, 50011, USA

    Andrew R. Korte & Young Jin Lee

  2. Ames Laboratory-USDOE, Ames, IA, 50011, USA

    Andrew R. Korte, Basil J. Nikolau & Young Jin Lee

  3. Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, 50011, USA

    Marna D. Yandeau-Nelson

  4. Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA, 50011, USA

    Basil J. Nikolau

  5. Center for Metabolic Biology, Iowa State University, Ames, IA, 50011, USA

    Basil J. Nikolau

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  1. Andrew R. Korte
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Correspondence to Young Jin Lee.

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Published in the topical collection Mass Spectrometry Imaging with guest editors Andreas Römpp and Uwe Karst.

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Korte, A.R., Yandeau-Nelson, M.D., Nikolau, B.J. et al. Subcellular-level resolution MALDI-MS imaging of maize leaf metabolites by MALDI-linear ion trap-Orbitrap mass spectrometer. Anal Bioanal Chem 407, 2301–2309 (2015). https://doi.org/10.1007/s00216-015-8460-5

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  • DOI: https://doi.org/10.1007/s00216-015-8460-5

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