Cole R, Matuszek G, See C, Rieder C L
Wadsworth Center for Laboratories and Research, Albany, New York 12201.
J Electron Microsc Tech. 1990 Oct;16(2):167-73. doi: 10.1002/jemt.1060160207.
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 LN2 creates an atmosphere below -120 degrees C in which the transfer can be accomplished.
本文介绍了一种简单且成本低廉的变速（1.0 - 5.8米/秒）气动无浸入式冷冻装置的详细设计。在涂有福尔马林的75目金查找器网格上生长的培养细胞，通过气动方式被驱动到由液氮（LN₂）冷却的丙烷/异戊烷（3:1）搅拌混合物中。通过一根入口管插入一个抽成部分真空的隔热箱，可防止样品在低温液体上方的低温蒸汽中过早冻结。真空将低温蒸汽吸入隔热箱中低温液体的液面高度，从而防止在低温液体上方积聚一层冷气。为防止样品在从低温液体转移到置换介质的过程中解冻，打开箱盖，通过一根贯穿箱体长度的波纹管强制通入压缩空气。由此产生的沸腾液氮会营造出低于 -120摄氏度的气氛，在此气氛中即可完成转移。
