Handley D A, Alexander J T, Chien S
J Microsc. 1981 Mar;121(Pt 3):273-82. doi: 10.1111/j.1365-2818.1981.tb01224.x.
The detailed design of a simple device for rapid quench-freezing of biological samples under reproducible conditions is presented. With spring-augmented descent, sample immersion velocity of 10 m s-1 into a cryogenic liquid is achieved. Biological samples, loaded in Balzers planchets, Denton holders, or a newly designed 'titanium envelope', are suitable for rapid-freezing with this device. Using 4 micrometers titanium foil, light weight (1 mg) streamlined holders can easily be made to enclose cell suspensions or tissue samples. The foil envelope is designed for efficient heat dissipation while protecting the sample from possible impact or flow distortions occurring from spring-augmented immersion. Human erythrocytes, quench-frozen in the titanium envelope, were prepared for electron microscopy by the freeze-substitution technique. Two opposing 25--30 micrometers surface zones were frozen in the apparent absence of ice. The extended depth of cryofixation is attributed to the advantages of thin foil in the titanium envelope design and the use of rapid-immersion technique.
本文介绍了一种简单装置的详细设计，该装置可在可重复的条件下对生物样品进行快速无淬灭冷冻。通过弹簧增强下降，可实现样品以10米/秒的速度浸入低温液体中。装载在Balzers平板、Denton固定器或新设计的“钛包膜”中的生物样品适用于该装置的快速冷冻。使用4微米厚的钛箔，可以轻松制作出重量轻（1毫克）的流线型固定器，用于包裹细胞悬液或组织样品。箔包膜的设计旨在实现高效散热，同时保护样品免受弹簧增强浸入过程中可能出现的冲击或流动变形的影响。通过冷冻替代技术制备了在钛包膜中淬灭冷冻的人类红细胞用于电子显微镜观察。在明显没有冰的情况下，两个相对的25 - 30微米表面区域被冷冻。冷冻固定深度的增加归因于钛包膜设计中薄箔的优点以及快速浸入技术的使用。
