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Cryo-SEM and TEM of High Pressure Frozen Cells - Some Technical Contributions

Published online by Cambridge University Press:  02 July 2020

Paul Walther
Paul Walther*
Affiliation:
Central Laboratory for Electron Microscopy, University of Ulm, D-89069, Ulm, Germanyhttp://www.uni-ulm.de/elektronenmikroskopie/http://www.nice.org.uk/page.aspx?o=43210
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Abstract

Imaging of fast frozen samples is the most direct approach for electron microscopy of biological specimen in a defined physiological state. It prevents chemical fixation and drying artifacts. High pressure freezing allows for ice-crystal-free cryo-fixation of tissue pieces up to a thickness of 200 urn and a diameter of 2 mm without prefixation. Such a frozen disc, however, is not directly amenable to electron microscopic observation: The structures of interest have to be made amenable to the electron beam, and the structures of interest must produce enough contrast to be recognized in the electron microscope. This can be achieved by freeze fracturing, cryo-sectioning or freeze substitution.

The figures show high pressure frozen bakers yeast saccharomyces cerevisiae in the cryo-SEM (Figures 1 and 2) and after freeze substitution in the TEM (Figure 3). For high pressure freezing either a Bal-Tec HPM 010 (Princ. of Liechtenstein; Figures 1 and 2), or a Wohlwend HPF (Wohlwend GmbH, Sennwald, Switzerland; Figure 3) were used.

Type
Cryoimmobilization, Freeze Substitution and Cryoem (Organized by S. Erlandsen)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 728 - 729
Copyright
Copyright © Microscopy Society of America 2001

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References

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