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Open AccessArticle

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

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.
Materials 2014, 7(4), 3160-3175; https://doi.org/10.3390/ma7043160
Received: 5 February 2014 / Revised: 10 March 2014 / Accepted: 11 April 2014 / Published: 21 April 2014
(This article belongs to the Section Biomaterials)
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. View Full-Text
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
MDPI and ACS Style

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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