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High-Pressure Freezing: Current State and Future Prospects

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Electron Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1117))

Abstract

In this chapter we discuss the latest developments in the field of high-pressure freezing (HPF). The Leica HPF machine EM HPM100 is discussed in detail due to significant changes compared to its predecessor model. Its centerpiece is a multipart polymer cartridge which holds the specimen carrier sandwich and guides it automatically through the freezing process until immersed in liquid nitrogen. The cartridge can be adapted to the specimen and carrier geometry to optimize the flow of liquid nitrogen and hence rapid cooling. Dedicated cartridges are available for a variety of different carriers, including carriers for samples of up to 5 mm in diameter. Cartridge-specific handling and carrier assemblies are described extensively for freezing samples in aluminum specimen carriers, cell cultures grown on Sapphire discs, suspensions for freeze-fracturing, and specimens for cryo-sectioning. Additionally, we include an advanced technique to freeze monolayer cell cultures on Sapphire discs with the Leica EM PACT2 HPF machine using a composite carrier.

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Acknowledgements

We would like to thank all colleagues in the field for fruitful and stimulating discussions and input, which were and still are indispensable for improving the application and technique of HPF. Special thanks go to Kim Rensing for sharing his experience and developed methods and for proofreading the manuscript. We wish also to thank all our collaborators for providing so many different and challenging samples.

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Authors and Affiliations

  1. Center for Microscopy and Image Analysis, University of Zurich, Zurich, Switzerland

    Andres Kaech & Urs Ziegler

Authors
  1. Andres Kaech
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  2. Urs Ziegler
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Editors and Affiliations

  1. University of Western Australia, Crawley, Australia

    John Kuo

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Kaech, A., Ziegler, U. (2014). High-Pressure Freezing: Current State and Future Prospects. 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_8

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  • DOI: https://doi.org/10.1007/978-1-62703-776-1_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-775-4

  • Online ISBN: 978-1-62703-776-1

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