Skip to main content

Botanical X-Ray Microanalysis in Cryoscanning Electron Microscopy

  • Protocol
Electron Microscopy

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 369))

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Chandler, J. A. (1977) X-ray Microanalysis in the Electron Microscope. North-Holland, Amsterdam.

    Google Scholar 

  2. Goldstein, J. I., Newbury, D. E., Echlin, P., et al. (2003) Scanning Electron Microscopy and X-ray Microanalysis. Kluwer Academic and Plenum, New York.

    Google Scholar 

  3. 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.

    Book  Google Scholar 

  4. Warley, A. (1997) X-ray microanalysis for biologists. Practical Methods in Electron Microscopy, vol. 16 (Glauert, A. M., ed.), Portland Press, London and Miami.

    Google Scholar 

  5. Echlin, P. (1992) Low-Temperature Microscopy and Analysis. Plenum Press, New York.

    Google Scholar 

  6. 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.

    Google Scholar 

  7. 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.

    Google Scholar 

  8. Brunner, I. and Frey, B. (2000) Detection and localization of aluminium and heavy metals in ectomycorrhizal Norway spruce seedlings. Environ. Poll. 108, 121–128.

    Article  CAS  Google Scholar 

  9. 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.

    Article  CAS  Google Scholar 

  10. 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.

    Article  CAS  Google Scholar 

  11. 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.

    Article  CAS  Google Scholar 

  12. 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.

    Article  Google Scholar 

  13. Zierold, K. (1988) X-ray microanalysis of freeze-dried and frozen-hydrated cryosections. J. Electron Microsc. Tech. 9, 65–82.

    Article  CAS  PubMed  Google Scholar 

  14. Marshall, A. T. (1998) X-ray microanalysis of frozen-hydrated biological bulk samples. Mikrochim. Acta Suppl. 15, 273–282.

    CAS  Google Scholar 

  15. Roomans, G. M. (1988) Quantitative X-ray microanalysis in biology. J. Electron Microsc. Tech. 9, 19–43.

    Article  CAS  PubMed  Google Scholar 

  16. 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.

    Google Scholar 

  17. Zierold, K. (2002) Limitations and prospects of biological electron probe X-ray microanalysis. J. Trace Microprobe Tech. 20, 181–196.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

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

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-294-6_26

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-573-6

  • Online ISBN: 978-1-59745-294-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics