Abstract:

The formation of intracellular ice of cells can cause severe cellular damage leading to many problems in cryopreservation. The broad bean was used as the research object, and the cytoskeleton was dissolved by cytochalasin B, and the freezing experiment was carried out at different cooling rates using a low temperature microscopic system. The experimental results show that the cells treated with cytochalasin B have higher crystallization temperature and shorter crystallization time during the freezing process, but the cytoskeleton has little effect on the growth process of intracellular ice. The external conditions play a key role, and the inoculation of ice crystals affects the formation temperature of ice crystals in the cells and the growth rate of ice crystals. Finally, the degree of damage to the cells was analyzed by light intensity map. It was found that the growth process of intracellular ice without cytoskeleton was similar to that of normal cells, but the crystallization rate was different, indicating that the cytoskeleton inside the cells affected the initiation of intracellular ice. Therefore, keeping the cytoskeleton intact before cryopreserving the plant sample can reduce the formation of intracellular ice and maintain cell morphology after rewarming.

Key words: microscopic measurement, microfilament skeleton, cytochalasin B, mass transfer, cell biology, interface

摘要：

细胞胞内冰的形成会导致严重的细胞损伤从而导致低温贮存中的诸多问题。以蚕豆为研究对象，用细胞松弛素B溶解细胞骨架，使用低温显微系统在不同的冷却速率下进行冷冻实验。实验结果表明，使用细胞松弛素B处理过的细胞在冷冻过程中结晶温度更高，结晶时间更短，但细胞骨架对胞内冰的生长过程影响较小。外界条件起着关键作用，接种冰晶影响细胞内冰晶的形成温度及冰晶的生长速率。最后，通过光强度图对细胞的损伤程度进行了分析。

关键词: 显微测量, 微丝骨架, 细胞松弛素B, 传质, 细胞生物学, 界面