一种简单快速植物组织冰冻切片方法

A Simple and Rapid Cryo-sectioning Method in Plant Tissue

比较不同冷冻方法对植物细胞超微结构的影响,结果表明:直接包埋法处理的植物细胞超微结构保存较好,而液氮冷冻处理的植物细胞内膜系统损伤严重。建立了一种直接包埋冷冻和适当回温相结合的方法,不仅可以制作出植物细胞基本结构保存完整的组织切片,而且避免了使用冰冻保护剂的弊端。其操作程序是:样品固定→冰冻与包埋→适当回温→快速切片→展片→染色。此法制作的切片可进行不同的染色和组织细胞化学测定,具有操作简便,易于推广的特点。

The effects of different freezing treatments on uttrastructure of plant cells during cryo-sectioning were compared. The results shown that the cell ultrastructure was well preserved with direct cryoembed method, while the endomembrane system in the cell was injured severely with liquid nitrogen cryopreservation. We proposed a method combining with cryoembed and modest thawing, by which not only could get integrated sections with the basic structure preserved, but also could avoid the abuse of using cyro-protected reagent. The procedure consisted of several steps as follows： fixing → refrigeration and embedding → modest thawing → rapid sectioning section flatting→ staining. The sections made by direct cryoembed with modest thawing could be stained and chemical determined in different ways. Besides, the characteristic of this procedure is simple and easy to be promoted.