SREL Reprint #2980

Cryoetch And Cryo-planing For Low Temperature HRSEM: SE-I Imaging Of Hydrated Multicellular, Microbial And Bioorganic Systems

R. P. Apkarian1, S. A. Shamsi2, S. A. Rizvi2, G. Benian3, A. L. Neal4, J. V. Taylor1, and S. N. Dublin1

1Integrated Microscopy & Microanalytical Facility, Emory University, Atlanta, GA 30322
2Department of Chemistry, Georgia State University, Atlanta, GA 30302
3Department of Pathology, Emory University School of Medicine, Atlanta, GA 30322
4Savannah River Ecology Laboratory, University of Georgia, Aiken, SC 29802

Introduction: Low temperature (LT) preparation of organic species and biological specimens in small volumes (< 5 µl) by plunging in liquefied ethane or high pressure freezing (HPF) ensures rapid cryoimmobilization for subsequent processing and imaging with cryo-HRSEM. Provided that specimen concentrations are high enough to act as its own cryoprotectant, hexagonal ice formation may be avoided and the sample is preserved in the low temperature state. Subsequent processing by cryogenic fracture, cryo-planing, and high vacuum sublimation (etching) relieve surfaces for structural analysis in the nanometer range. Successful LT processing is completed by application of a thin monoatomic chromium (Cr) film that enriches the SE-I contrast for high resolution cryo-HRSEM recordings.

SREL Reprint #2980

Apkarian, R. P., S. A. Shamsi, S. A. Rizvi, G. Benian, A. L. Neal, J. V. Taylor, and S. N. Dublin. 2006. Cryoetch And Cryo-planing For Low Temperature HRSEM: SE-I Imaging Of Hydrated Multicellular, Microbial And Bioorganic Systems. Microscopy and Microanalysis 12(Supp 2): 1120-1121.

This information was provided by the University of Georgia's Savannah River Ecology Laboratory (srel.uga.edu).