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Refining Equipment for High Resolution in-Lens Cryo-Sem

Published online by Cambridge University Press:  02 July 2020

Robert P. Apkarian ,
Stephen Lee  and
Jason Keiper
Robert P. Apkarian
Affiliation:
Integrated SEM & Microanalytical Facility, Department of Chemistry, Emory University, Atlanta, GA, 30322
Stephen Lee
Affiliation:
Integrated SEM & Microanalytical Facility, Department of Chemistry, Emory University, Atlanta, GA, 30322
Jason Keiper
Affiliation:
Integrated SEM & Microanalytical Facility, Department of Chemistry, Emory University, Atlanta, GA, 30322
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Extract

Advanced metal coating methods have greatly improved the localization of secondary electrons (SEI) from biological specimens staged in an in-lens field emission SEM. Using these metal coatings, molecular level structural information from cell membranes and isolated bio-molecules have been more accurately recorded from frozen-hydrated samples due to the presence of bound water rather than from chemically fixed and dried samples. Although in-lens cryo-SEM systems have provided images with equivalent resolution to TEM-replicas, several conditions are encountered which require optimizing techniques and hardware.

A drop of hydrated Bakers yeast on a gold Balzers carrier was plunge-frozen in ethane at its melting point. Thermal properties of ethane, such as the large difference between melting and boiling points, and its low viscosity increase the heat transfer coefficient during plunging thereby leading to faster cooling of the sample than with other cryogens. Carriers were transferred to the cryo-workstation of the Oxford CT 3500 system.

Type
New Trends in Scanning Electron Microscopy and Microanalysis
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 258 - 259
Copyright
Copyright © Microscopy Society of America

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References

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