Hunt C J
Cryobiology. 1984 Aug;21(4):385-402. doi: 10.1016/0011-2240(84)90077-4.
Recent studies have led to the conclusion that extracellular ice per se can damage whole organs and tissues. Thus information on the amount and distribution of ice is an important factor in the design of cooling regimens that avoid intracellular ice formation and attempt to localize the ice formed in areas of the tissue where its disruptive effects can be minimized. Furthermore, ultrastructural studies at subfreezing temperatures can enhance the interpretation of information gained from morphological and function studies conducted before cooling and after rewarming. Although many techniques exist for observing and recording structure in the frozen state, not all are applicable to tissues or organs. Freeze-substitution and isothermal freeze-fixation provide two flexible techniques to explore the frozen state. Isothermal freeze-fixation is most suitable for studies close to the melting point, while freeze-substitution can be used at lower temperatures, extending as far as -120 degrees C. A careful choice of technique can provide an accurate assessment of the amount and distribution of the ice phase and the structure of the tissue matrix.
最近的研究得出结论，细胞外冰本身会损害整个器官和组织。因此，关于冰的数量和分布的信息是设计冷却方案的一个重要因素，该方案要避免细胞内结冰，并试图将形成的冰定位在组织中其破坏作用可降至最低的区域。此外，在亚冰点温度下的超微结构研究可以增强对冷却前和复温后进行的形态学和功能研究所得信息的解读。虽然存在许多用于观察和记录冷冻状态下结构的技术，但并非所有技术都适用于组织或器官。冷冻置换和等温冷冻固定提供了两种灵活的技术来探索冷冻状态。等温冷冻固定最适合接近熔点的研究，而冷冻置换可用于较低温度，低至 -120 摄氏度。仔细选择技术可以准确评估冰相的数量和分布以及组织基质的结构。
