Fischer Andrew H, Jacobson Kenneth A, Rose Jack, Zeller Rolf
CSH Protoc. 2008 Aug 1;2008:pdb.prot4991. doi: 10.1101/pdb.prot4991.
INTRODUCTIONCryosections are rapidly and relatively easily prepared prior to fixation, and they provide a good system for visualizing fine details of the cell. Although cryosections are physically less stable than paraffin- or resin-embedded sections, they are generally superior for the preservation of antigenicity and therefore the detection of antigens by microscopy. The preparation of cryosections does not involve the dehydration steps typical of other sectioning methods, and, furthermore, sectioning, labeling, and observation of specimens can usually be carried out in one day. In general, the sample is frozen quickly in either isopentane or liquid nitrogen. (Small samples such as cells and small tissues may be mixed in a slurry of an inert support medium such as optimal cutting temperature [OCT] compound before freezing). Rapid freezing reduces ice crystal formation and minimizes morphological damage. Frozen sections may be used for a variety of procedures, including immunochemistry, enzymatic detection, and in situ hybridization. A protocol for cryosectioning is presented here.
引言
冷冻切片在固定前制备迅速且相对容易，为观察细胞的细微细节提供了良好的系统。尽管冷冻切片在物理稳定性上不如石蜡或树脂包埋切片，但它们在保存抗原性方面通常更具优势，因此更适合通过显微镜检测抗原。冷冻切片的制备不涉及其他切片方法典型的脱水步骤，此外，标本的切片、标记和观察通常可在一天内完成。一般来说，样本在异戊烷或液氮中快速冷冻。（细胞和小组织等小样本在冷冻前可与惰性支持介质如最佳切片温度[OCT]化合物的浆液混合）。快速冷冻可减少冰晶形成并将形态损伤降至最低。冷冻切片可用于多种操作，包括免疫化学、酶检测和原位杂交。本文介绍了冷冻切片的操作流程。
