Method and apparatus for vitrification of water or moisture-cont

Refrigeration – Storage of solidified or liquified gas – Including cryostat

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62 78, F25B 1900

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active

054938651

ABSTRACT:
To vitrify a sample, for example a biological sample which has a high water content, without the formation of ice crystals, the sample (10) is placed in a closed test chamber (11) having a chamber outlet (35). The chamber (11), with the sample therein, is filled with a primary fluid without cooling the sample to freezing temperature. This primary fluid may be an alcohol. The sample is vitrified by applying a high-pressure cryogenic fluid against the sample at a substantial pressure. The step of maintaining the pressure is accomplished, in accordance with the invention, by maintaining the outlet (35) from the chamber (11) at least essentially closed by a closing element, typically a valve (41), until the pressure within the sample chamber (11) has reached a predetermined value. The primary fluid may be a portion of the cryogenic fluid itself, for example heated, or even cooled, to an appropriate temperature which, however, will not cause the sample to freeze. Automatic interlocks (63; 32) can be provided to ensure that the high-pressure cryogenic fluid is applied only after the chamber has been filled and pressurized by the primary fluid. A pressure rise to 2,100 bar, and more, in 20 milliseconds can be obtaining, resulting in a high cooling rate of at least 20,000.degree. C. per second, which cools the sample. Liquid nitrogen is a suitable cryogenic fluid.

REFERENCES:
patent: 4559298 (1985-12-01), Fahy
patent: 4685305 (1987-08-01), Burg
patent: 4688387 (1987-08-01), Conaway
patent: 4753887 (1988-06-01), Bellare et al.
patent: 4799361 (1989-01-01), Linner
Publication by BALZERS Union AG of Balzers, Principality of Liechtenstein: "Elektronenmikroskopie--Hochdruck-Gefriermaschine HPM 010" ( Electron Microscopy--High-Pressure Freezing Machine HPM 010).
Studer et al, High Pressure Freezing comes of Age, Scanning Microscopy, Supplement 3, 1989, pp. 253-269.
Kanno et al. (1975), "Supercooling of water to -92.degree. C. under pressure". Science 189:880-881.
Moor H. (1987), Theory and practice of high pressure freezing. In: Steinbrecht R. A. Zierold K (eds) Cryotechniques in Biological Electron Microscopy, Berlin, Springer, 175-191.
Michel et al. (1991), J. Microsc. 163:3-18.

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