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Preservation of Biomembranes by High Pressure Freezing ?

Published online by Cambridge University Press:  02 July 2020

Martin Müller ,
Jens Listemann ,
Eyal Shimony  and
Paul Walther
Martin Müller
Affiliation:
Laboratory for Electron Microscopy, ETH-Zürich, CH-8092, Zürich, Switzerland
Jens Listemann
Affiliation:
Laboratory for Electron Microscopy, ETH-Zürich, CH-8092, Zürich, Switzerland
Eyal Shimony
Affiliation:
Laboratory for Electron Microscopy, ETH-Zürich, CH-8092, Zürich, Switzerland
Paul Walther
Affiliation:
Laboratory for Electron Microscopy, ETH-Zürich, CH-8092, Zürich, Switzerland
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Extract

Sample preparation techniques for electron-microscopy (EM) dictate our perception of the microworld: any structural information which is lost or distorted during preparation can not be regenerated later and might lead to wrong interpretation of the observed micrograph.

Cryofixation based procedures for specimen preparation can avoid most of the structural alterations associated with standard techniques based on chemical fixation. The ultrastructure can be represented in a near “native state” thanks to the high time resolution for dynamic cellular events (1).

High pressure freezing (2) permits to cryoimmobilize biological samples up to approx. 200μm thick, in contrast to rapid freezing procedures at ambient pressure that are useful to cryoimmobilize samples up to 10-20 μm thick. The actual samplethickness that can be adequately frozen (=without visible damage due to ice crystal formation in freeze-substituted or freeze-fractured specimens) depends on the concentration of naturally occuring substances that exhibit cryoprotective activities.

Type
Cryotechniques, Immunocytochemistry, and Electron Microscopy II. Cells and Tissues
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 428 - 429
Copyright
Copyright © Microscopy Society of America

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References

REFERENCES

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