Carson J L, Collier A M, Smith C A
J Ultrastruct Res. 1984 Oct;89(1):23-33. doi: 10.1016/s0022-5320(84)80020-9.
Freshly isolated, viable hamster tracheal epithelium was rapidly frozen in a jet of liquid nitrogen-cooled propane followed by freeze-fracture, deep etching, and rotary shadowing. Examination of the replicas by transmission electron microscopy revealed characteristic features of both ciliated and nonciliated cells, profiles of tight junctional complexes, and two distinct types of membrane particle complexes. The findings of this study suggest that rapid freezing of viable biological specimens coupled with freeze-fracture, deep etching, and rotary shadowing may provide a useful approach to achieving some additional perspectives of cell structure and function. Since the experimental protocol avoided any use of chemical fixation and processing procedures generally employed in electron microscopy, this study also validates for airway epithelium the details of fine structure observed using more conventional ultrastructural methods.
新鲜分离的、有活力的仓鼠气管上皮细胞在液氮冷却的丙烷气流中迅速冷冻，随后进行冷冻断裂、深度蚀刻和旋转阴影投射。通过透射电子显微镜检查复制品，揭示了纤毛细胞和非纤毛细胞的特征、紧密连接复合体的轮廓以及两种不同类型的膜颗粒复合体。本研究结果表明，对有活力的生物标本进行快速冷冻，再结合冷冻断裂、深度蚀刻和旋转阴影投射，可能为深入了解细胞结构和功能提供一种有用的方法。由于实验方案避免了电子显微镜中通常使用的任何化学固定和处理程序，本研究还验证了使用更传统的超微结构方法观察到的气道上皮细胞精细结构的细节。
