A simple pneumatic device for plunge-freezing cells grown on electron microscopy grids.

A simple pneumatic device for plunge-freezing cells grown on electron microscopy grids.
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一种简单的气动装置,用于冷冻在电子显微镜网格上生长的细胞。

DOI:
10.1002/jemt.1060160207
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发表时间:
1990
期刊:
Journal of electron microscopy technique
影响因子:
--
通讯作者:
Rieder,CL
Rieder,CL
中科院分区:
--
文献类型:
--
作者:
Cole,R;Matuszek,G;See,C;Rieder,CL

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介绍了一种简单而廉价的变速(1.0-5.8 m s−1)气动插入式冷冻装置的详细设计。将在Formvar涂覆的75目黄金网格上生长的培养细胞快速驱动到由液氮(LN 2)冷却的丙烷/异戊烷(3:1)的搅拌混合物中。样品在冷冻剂上方的低温蒸气中过早冻结是通过将样品通过入口管插入到施加部分真空的绝缘箱中来防止的。低温蒸汽通过真空在液体制冷剂的水平处被吸入箱中,从而防止冷气层聚集在制冷剂上方。为了防止样品在从冷冻剂转移到置换介质的过程中解冻,将箱顶移除,并迫使压缩空气通过沿箱长度延伸的波纹管。由此产生的沸腾的液氮2创造了一个低于−120°C的气氛,在其中可以完成转移。
A detailed design for a simple and inexpensive variable‐speed (1.0–5.8 m s−1) pneumatic plunge‐freezing device is presented. Cultured cells, grown on Formvar‐coated 75‐mesh gold finder grids, are pneumatically driven into a stirring mixture of propane/isopentane (3:1) cooled by liquid nitrogen (LN2). Premature freezing of the sample in the cryogenic vapors above the cryogen is prevented by plunging through an entry tube into an insulating box, to which a partial vacuum is applied. The cryogenic vapors are drafted into the box at the level of the liquid cryogen by the vacuum, thereby preventing a layer of cold gas from collecting above the cryogen. To prevent the sample from thawing during transfer from the cryogen to the substitution medium, the box top is removed and compressed air is forced through a corrugated tube running the length of the box. The resulting boiling LN2creates an atmosphere below −120°C in which the transfer can be accomplished.
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