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Modifications Needed for Performing High-Resolution Cryosem in a Hitachi S-4700 Fesem Using an Emitech K1250 Cryo-Preparation System

Published online by Cambridge University Press:  02 July 2020

Ya Chen ,
Chris Frethem  and
Stanley L. Erlandsen
Ya Chen
Affiliation:
EM Facility, Department of Genetics, Cell Biology, and Development University of Minnesota-Medical School, Minneapolis, MN, 55455, USA
Chris Frethem
Affiliation:
EM Facility, Department of Genetics, Cell Biology, and Development University of Minnesota-Medical School, Minneapolis, MN, 55455, USA
Stanley L. Erlandsen
Affiliation:
EM Facility, Department of Genetics, Cell Biology, and Development University of Minnesota-Medical School, Minneapolis, MN, 55455, USA
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Abstract

Cryo-techniques have been successfully used in scanning electron microscopy (SEM). They are especially promising for high-resolution SEM to improve specimen preservation and reduce radiation damage [1, 2]. A number of cryo-preparation systems are commercially available for SEM, however, our experience has shown that modifications are needed to perform highresolution imaging (>50,000x).

Emitech K1250 system consists of a sample preparation chamber, control unit, and cryo-stage. Magnetron sputter coating is standard and electron-beam evaporation is optional. A vacuum transfer device facilitates the sample transfer between the preparation chamber and the SEM to prevent contamination. The Emitech cryo-stage replaces the Hitachi S-4700 standard stage and the cryo-stage temperature is monitored and controlled by the Emitech control unit.

The specimen is mounted on a sample holder that mounts to the cryo-stage. Therefore, the distance from the specimen to cryo-stage, the thermal capacity of the sample holder, and the thermal contact between them will affect the actual temperature of the specimen.

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. 720 - 721
Copyright
Copyright © Microscopy Society of America 2001

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References

1. Erlandsen, S. L., Frethem, C., Chen, Y.J. Histotechnology 23 (2000) 249–259.CrossRefGoogle Scholar

2. Chen, Y., Centonze, V. E., Verkhovsky, A., Borisy, G. G.J. Microsc. 179 (1995) 67–76CrossRefGoogle Scholar

3. P., Walther, Wehrli, E., Hermann, R., Miiller, M.J. Microsc. 179 (1995) 229–237.Google Scholar