Abstract
Modern microscopy in plant sciences has evolved in the direction of providing ultra-structural and analytical information simultaneously. Energy-dispersive X-ray microanalysis (EDX) is a powerful technique that allows the qualitative and quantitative measurement of many elements of physiological interest at the cellular and subcellular level. The most significant advance has been the development of freezing techniques to study cells in plant tissues by EDX in the cryoscanning electron microscopy. Cryofixation is fast enough to retain the original distributions of inorganic elements of tissue electrolytes sufficiently for microanalytical studies. This approach may have broad application for various types of localizations of relevance to plant physiology, environmental pollution, and plant-microbe interactions. In this chapter, the experimental procedure of analytical cryoscanning electron microscopy applied to botanical samples is outlined.
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References
Chandler, J. A. (1977) X-ray Microanalysis in the Electron Microscope. North-Holland, Amsterdam.
Goldstein, J. I., Newbury, D. E., Echlin, P., et al. (2003) Scanning Electron Microscopy and X-ray Microanalysis. Kluwer Academic and Plenum, New York.
Sigee, D. C., Morgan, A. J., Sumner, A. T., and Warley, A. (1993) X-ray Microanalysis in Biology: Experimental Techniques and Applications. Cambridge University Press, Cambridge.
Warley, A. (1997) X-ray microanalysis for biologists. Practical Methods in Electron Microscopy, vol. 16 (Glauert, A. M., ed.), Portland Press, London and Miami.
Echlin, P. (1992) Low-Temperature Microscopy and Analysis. Plenum Press, New York.
Jeffree, C. E. and Read, N. D. (1991) Ambient-and low-temperature scanning electron microscopy, in Electron Microscopy of Plant Cells, (Hall, J. L. and Hawes, C., eds.), Academic Press, London, pp. 313–413.
Zierold, K. and Steinbrecht, R. A. (1987) Cryofixation of diffusible elements in cells and tissues for electron probe microanalysis, in Cryotechniques in Biological Electron Microscopy (Steinbrecht, R. A. and Zierold, K., eds.), Springer, Berlin, Heidelberg, New York, pp. 272–282.
Brunner, I. and Frey, B. (2000) Detection and localization of aluminium and heavy metals in ectomycorrhizal Norway spruce seedlings. Environ. Poll. 108, 121–128.
Frey, B., Brunner, I., Walther, P., Scheidegger, C., and Zierold, K. (1997) Element localization in ultrathin cryosections of high-pressure frozen ectomycorrhizal spruce roots. Plant Cell Environ. 20, 929–937.
Frey, B., Keller, C., Zierold, K., and Schulin, R. (2000) Distribution of Zn in functionally different leaf epidermal cells of the hyperaccumulator Thlaspi caerulescens. Plant Cell Environ. 23, 675–687.
Frey, B., Zierold, K., and Brunner, I. (2000) Extracellular complexation of Cd in the Hartig net and cytosolic Zn sequestration in the fungal mantle of Picea abies-Hebeloma crustuliniforme ectomycorrhizas. Plant Cell Environ. 23, 1257–1265.
Stelzer, R. and Lehmann, H. (1993) Recent developments in electron microscopical techniques for studying ion localization in plant cells. Plant Soil 155/156, 33–43.
Zierold, K. (1988) X-ray microanalysis of freeze-dried and frozen-hydrated cryosections. J. Electron Microsc. Tech. 9, 65–82.
Marshall, A. T. (1998) X-ray microanalysis of frozen-hydrated biological bulk samples. Mikrochim. Acta Suppl. 15, 273–282.
Roomans, G. M. (1988) Quantitative X-ray microanalysis in biology. J. Electron Microsc. Tech. 9, 19–43.
Van Steveninck, R. F. M. and Van Steveninck, M. E. (1991) Microanalysi, in Electron Microscopy of Plant Cells, (Hall, J. L. and Hawes, C., eds.), Academic Press, London, pp. 415–455.
Zierold, K. (2002) Limitations and prospects of biological electron probe X-ray microanalysis. J. Trace Microprobe Tech. 20, 181–196.
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Frey, B. (2007). Botanical X-Ray Microanalysis in Cryoscanning Electron Microscopy. In: Kuo, J. (eds) Electron Microscopy. Methods in Molecular Biology™, vol 369. Humana Press. https://doi.org/10.1007/978-1-59745-294-6_26
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DOI: https://doi.org/10.1007/978-1-59745-294-6_26
Publisher Name: Humana Press
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