A new approach for cryofixation by high‐pressure freezing

A newly designed high‐pressure freezing machine for cryofixation was established and tested (Leica EMPACT), based on ideas originally proposed by Moor & Riehle in 1968. The new machine, essentially an improved version of our prototype, pressurizes the sample to 2000 bar in a small container (usi...

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Bibliographic Details
Published in:Journal of microscopy (Oxford) 2001-09, Vol.203 (3), p.285-294
Main Authors: Studer, D., Graber, W., Al‐Amoudi, A., Eggli, P.
Format: Article
Language:English
Subjects:
Animals
Atmospheric Pressure
Cryofixation
cryomicroscopy
cryosectioning
electron microscopy
Freeze Substitution - instrumentation
Freeze Substitution - methods
freeze‐substitution
Freezing
high‐pressure freezing
Liver - cytology
Liver - ultrastructure
Microscopy, Electron - methods
Microtomy
Peripheral Nerves - cytology
Peripheral Nerves - ultrastructure
Plant Leaves - cytology
Plant Leaves - ultrastructure
Rats
Temperature
Tissue Fixation - instrumentation
Tissue Fixation - methods
Yeasts - cytology
Yeasts - ultrastructure
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Summary:A newly designed high‐pressure freezing machine for cryofixation was established and tested (Leica EMPACT), based on ideas originally proposed by Moor & Riehle in 1968. The new machine, essentially an improved version of our prototype, pressurizes the sample to 2000 bar in a small container (using methylcyclohexane as hydraulic fluid) and at the same time cools the outer surface of the container with a jet of liquid nitrogen. The advantage of this approach is that the machine uses little liquid nitrogen and can be built small and light. The machine is able to vitrify and freeze well a variety of specimens, for example, plant leaves, yeast cells, liver or nerve tissue (more samples are shown at: http://www.ana.unibe.ch/empact). Cooling efficiency is the same as in the traditional machines that use liquid nitrogen to pressurize and simultaneously cool the sample.
ISSN:0022-2720
1365-2818
DOI:10.1046/j.1365-2818.2001.00919.x