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Improved cryofixation applicable to freeze etching.

L Bachmann, W W Schmitt

Proceedings of the National Academy of Sciences of the United States of America

|September 1, 1971

Summary

Spray-freeze-etching minimizes cryofixation artifacts in solute systems. This novel cryofixation technique offers superior results for studying biological samples without antifreeze.

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Area of Science:

Cryo-electron microscopy
Biophysics
Materials science

Background:

Standard cryofixation methods can introduce artifacts, such as segregation of solutes, during the freezing of biological and model systems.
These artifacts complicate the accurate structural analysis of dissolved or dispersed materials using techniques like freeze-etching.

Purpose of the Study:

To develop and evaluate a new cryofixation technique, spray-freeze-etching, to overcome the limitations of conventional freezing methods.
To assess the efficacy of spray-freeze-etching in reducing segregation artifacts in solute model systems.

Main Methods:

Developed a spray-freeze-etching technique involving spraying specimens into a liquid cryomedium.
Collected frozen droplets using butylbenzene at -85°C to form a composite specimen for freeze-etching.
Tested the method using 5% glycerol solutions as a model system.

Main Results:

Spray-freeze-etching significantly reduced segregation artifacts compared to standard freezing techniques.
The results achieved with spray-freeze-etching for glycerol solutions were comparable to those obtained using high-pressure freezing.
The technique enables the investigation of solute systems and the study of unicellular organisms or cellular fractions without antifreeze.

Conclusions:

Spray-freeze-etching is a highly effective cryofixation method for preserving the native structure of solute systems.
This technique significantly improves the quality of freeze-etched specimens, making freeze-etching a more reliable method for structural studies.
The ability to avoid antifreeze agents broadens the applicability of freeze-etching for delicate biological samples.

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