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Environmental scanning electron microscopy (ESEM)—a versatile tool in studying plants

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Abstract

Environmental scanning electron microscopy (ESEM) enables the investigation of hydrated and uncoated plant samples and the in situ observation of dynamic processes. Water vapor in the microscope chamber takes part in secondary electron detection and charge prevention. Two ESEM modes are available and offer a broad spectrum of applications. The environmental or wet mode prevents sample dehydration by the combination of sample cooling (5°C) and a vapor pressure of 4–6 Torr. In the low vacuum mode, the maximum chamber pressure is limited to 1 Torr (corresponding to about 5% relative humidity in the chamber) and allows the simultaneous use of a backscattered electron detector for imaging material contrast. A selection of characteristic plant samples and various applications are presented as a guide to ESEM for plant scientists. Leaf surfaces, trichomes, epicuticular waxes, and inorganic surface layers represent samples being comparatively resistant to dehydration, whereas callus cells and stigmatic tissue are examples for dehydration- and beam-sensitive samples. The potential of investigating dynamic processes in situ is demonstrated by studying anther opening, by tensile testing of leaves, and by performing hydration/dehydration experiments by changing the vapor pressure. Additionally, automated block-face imaging and serial sectioning using in situ ultramicrotomy is presented. The strengths and weaknesses of ESEM are discussed and it is shown that ESEM is a versatile tool in plant science.

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Acknowledgements

Their work and support was very helpful and we want to thank Regina Willfurth for her help in the sample preparation, Alexandra Jammer for providing the callus samples, Günther Zellnig for providing the imbedded Pelargonium sample for in situ ultramicrotomy, and Helga Hammer for her excellent work in the greenhouse. The constructive criticism of the reviewers is greatly acknowledged.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Plant Sciences, University of Graz, Schubertstraße 51, 8010, Graz, Austria

    Edith Stabentheiner

  2. Institute for Electron Microscopy, Graz University of Technology and Center for Electron Microscopy, Steyrergasse 17, 8010, Graz, Austria

    Armin Zankel & Peter Pölt

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  1. Edith Stabentheiner
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  2. Armin Zankel
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  3. Peter Pölt
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Correspondence to Edith Stabentheiner.

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Stabentheiner, E., Zankel, A. & Pölt, P. Environmental scanning electron microscopy (ESEM)—a versatile tool in studying plants. Protoplasma 246, 89–99 (2010). https://doi.org/10.1007/s00709-010-0155-3

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  • DOI: https://doi.org/10.1007/s00709-010-0155-3

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