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Materials 2014, 7(4), 3160-3175; doi:10.3390/ma7043160
Article

Quantitative X-ray Elemental Imaging in Plant Materials at the Subcellular Level with a Transmission Electron Microscope: Applications and Limitations

1
,
2
,
2
 and
2,*
1 College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China 2 Forstbotanik und Baumphysiologie, Büsgen-Institute, Georg-August Universität Göttingen, Göttingen 37077, Germany
* Author to whom correspondence should be addressed.
Received: 5 February 2014 / Revised: 10 March 2014 / Accepted: 11 April 2014 / Published: 21 April 2014
(This article belongs to the Section Biomaterials)
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Abstract

Energy-dispersive X-ray microanalysis (EDX) is a technique for determining the distribution of elements in various materials. Here, we report a protocol for high-spatial-resolution X-ray elemental imaging and quantification in plant tissues at subcellular levels with a scanning transmission electron microscope (STEM). Calibration standards were established by producing agar blocks loaded with increasing KCl or NaCl concentrations. TEM-EDX images showed that the salts were evenly distributed in the agar matrix, but tended to aggregate at high concentrations. The mean intensities of K+, Cl, and Na+ derived from elemental images were linearly correlated to the concentrations of these elements in the agar, over the entire concentration range tested (R > 0.916). We applied this method to plant root tissues. X-ray images were acquired at an actual resolution of 50 nm ´ 50 nm to 100 nm ´ 100 nm. We found that cell walls exhibited higher elemental concentrations than vacuoles. Plants exposed to salt stress showed dramatic accumulation of Na+ and Cl in the transport tissues, and reached levels similar to those applied in the external solution (300 mM). The advantage of TEM-EDX mapping was the high-spatial-resolution achieved for imaging elemental distributions in a particular area with simultaneous quantitative analyses of multiple target elements.
Keywords: TEM; EDX; agar standard; Populus euphratica; root; cortex; cell wall; vacuole; xylem vessel; salt tolerance TEM; EDX; agar standard; Populus euphratica; root; cortex; cell wall; vacuole; xylem vessel; salt tolerance
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Chen, S.; Diekmann, H.; Janz, D.; Polle, A. Quantitative X-ray Elemental Imaging in Plant Materials at the Subcellular Level with a Transmission Electron Microscope: Applications and Limitations. Materials 2014, 7, 3160-3175.

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