Abstract
This chapter describes protocols using formalin-acetic acid-alcohol (FAA) to fix plant tissues for studying biomineralization by means of scanning electron microscopy (SEM) and qualitative energy-dispersive X-ray microanalysis (EDX). Specimen preparation protocols for SEM and EDX mainly include fixation, dehydration, critical point drying (CPD), mounting, and coating. Gold-coated specimens are used for SEM imaging, while gold- and carbon-coated specimens are prepared for qualitative X-ray microanalyses separately to obtain complementary information on the elemental compositions of biominerals. During the specimen preparation procedure for SEM, some biominerals may be dislodged or scattered, making it difficult to determine their accurate locations, and light microscopy is used to complement SEM studies. Specimen preparation protocols for light microscopy generally include fixation, dehydration, infiltration and embedding with resin, microtome sectioning, and staining. In addition, microwave processing methods are adopted here to speed up the specimen preparation process for both SEM and light microscopy.
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Acknowledgements
This work was supported by the Australian Research Council, Mineral and Energy Research Institute of Western Australia, Newcrest Mining Ltd. (Telfer Gold Mine), the School of Plant Biology and the Centre for Microscopy, Characterisation and Analysis, The University of Western Australia. We thank Chris Bray who provided the Olympus BX43 microscope. Prof. Hans Lambers is acknowledged for reviewing the draft of this chapter.
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He, H., Kirilak, Y. (2014). Application of SEM and EDX in Studying Biomineralization in Plant Tissues. In: Kuo, J. (eds) Electron Microscopy. Methods in Molecular Biology, vol 1117. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-776-1_29
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DOI: https://doi.org/10.1007/978-1-62703-776-1_29
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