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Published on June 20, 2008; 10.1104/pp.107.109215

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Received September 14, 2007
Accepted May 26, 2008

Imaging Nutrient Distributions in Plant Tissue using Time of Flight Secondary Ion Mass Spectrometry and Scanning Electron Microscopy

Ralf Metzner , Heike Ursula Schneider , Uwe Breuer , and Walter Heinz Schroeder *

Central Division of Analytical Chemistry, 2 Phytosphere Institute (ICG-3), Research Center Julich, 52425 Julich, Germany

A new approach to trace the transport routes of macronutrients in plants at the level of cells and tissues, and to measure their elemental distributions, was developed for investigating the dynamics and structure–function relations of transport processes. Stem samples from Phaseolus vulgaris were used as a test system. Shock-freezing and cryo-preparation were combined in a cryogenic chain with cryo-time-of-flight secondary ion mass spectrometry (cryo-ToF-SIMS) for element and isotope specific imaging. Cryo scanning electron microscopy (cryo-SEM) was integrated into the cryogenic workflow to assess the qualities of structural preservation. We evaluated the capability of these techniques to monitor transport pathways and processes in xylem and associated tissues using supplementary sodium and tracers for potassium, rubidium and 41K, added to the transpiration stream. Cryo-ToF-SIMS imaging produced detailed mappings of water, potassium, calcium, magnesium, the potassium tracers and sodium without quantification. Lateral resolutions ranged from 10 µm in survey mappings and high mass resolution to ca. 1 µm in high-lateral resolution imaging in reduced areas and lower mass resolution. The tracers Rb and 41K as well as Na were imaged with high sensitivity in xylem vessels and surrounding tissues. The isotope signature of the stable isotope tracer was utilized for relative quantification of the 41K tracer as fraction of total potassium at the single pixel level. Cryo-SEM confirmed that tissue structures had been preserved with sub cellular detail throughout all procedures. Overlays of cryo-ToF-SIMS images onto the corresponding SEM images allowed detailed correlation of nutrient images with sub-cellular structures.






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