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Freezing Techniques: History, Comparisons, and Applications

Published online by Cambridge University Press:  14 March 2018

Bill Graham ,
Jotham R. Austin II ,
Andres Kaech  and
John E. Heuser
Bill Graham*
Affiliation:
Bibst Labs Brookline, NH
Jotham R. Austin II
Affiliation:
University of Chicago Chicago, IL
Andres Kaech
Affiliation:
University of Zurich, Switzerland
John E. Heuser
Affiliation:
Washington Univ. School of Med., St. Louis, MO
*
bill@bibst.com

Extract

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Specimen preparation techniques have evolved hand in hand with microscopy since the first microscopes. Since the introduction of the first Electron Microscope (EM) in the 1930’s, the basic problem with biological electron microscopy has been to preserve the structure of soft condensed, hydrated matter (e.g. tissues, cells, proteins, etc.) so that they can be viewed in the harsh environment of the electron microscope's high vacuum and ionizing radiation. For this, cells must be “fixed” with chemical cross-linkers, commonly glutaraldehyde, formaldehyde or some combination of both, stained with heavy metals (osmium tetroxide that provides contrast of biological components), dehydrated with an organic solvent, and infiltrated with a resin for eventual thin-sectioning. Only then can it be viewed with the EM. Such treatment with chemical fixatives and stains remains the standard approaches to arrest biological processes in cells or tissues, but at the cost of introducing clearly recognizable artifacts.

Type
Research Article
Information
Microscopy Today , Volume 16 , Issue 5 , September 2008 , pp. 12 - 17
Copyright
Copyright © Microscopy Society of America 2008

References

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