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A Versatile High-Vacuum Cryo-Transfer for Cryo-FESEM, Cryo-SPM and other Imaging Techniques

Published online by Cambridge University Press:  02 July 2020

Martin Ritter ,
Didier Henry ,
Stefan Wiesner ,
Stephan Pfeiffer  and
Roger Wepf
Martin Ritter
Affiliation:
Bal-Tec AG, FL -9496 Bakers
Didier Henry
Affiliation:
HPI, University Clinic Eppendorf, Martinistr.52, D- 20251, Hamburg;
Stefan Wiesner
Affiliation:
Beiersdorf AG, Analytical Microscopy Unit, Unnastr. 48, D-20253, Hamburg;
Stephan Pfeiffer
Affiliation:
Beiersdorf AG, Analytical Microscopy Unit, Unnastr. 48, D-20253, Hamburg;
Roger Wepf
Affiliation:
Beiersdorf AG, Analytical Microscopy Unit, Unnastr. 48, D-20253, Hamburg;
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Extract

A structure preservation of biological and organic samples, close to native state, can only be reached by cryo immobilization techniques. Cryo immobilization allows not only to preserve the high structural integrity but also to arrest dynamic processes in the μs- ms range.

After freezing the sample and preparing the surface of interest, it is important to prevent the sample from ice crystal damage, removal of structural water, condensation of water or other contaminants until imaging. Therefore, ideally the samples are kept below the recrystallisation temperature of water (< 147K) during the transfer from the preparation environment into the imaging chamber.

For the transfer of frozen samples several concepts have been followed in the,past: a) the specimen after manipulation/preparation were submersed in liquid nitrogen and transferred to the cold stage of the microscope or b) a preparation chamber was permanently attached to the microscope column allowing the direct transfers between the preparation chamber and the cold stage in the microscope. These concepts allow either a high grade of flexibility combined with a high risk of contamination or to prevent contamination but combined with inflexibility. In addition the later also does not allow using the microscope during the specimen preparation procedure, nor transferring the specimen to an other imaging device.

Type
Cryotechniques, Immunocytochemistry, and Electron Microscopy I. Molecular Approach
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. 424 - 425
Copyright
Copyright © Microscopy Society of America

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