冰冻切片技术在铁皮石斛显微结构上的应用

Cryo-sectioning Technique for Microstructure Studies on Dendrobium officinale

为了快速高效的观察兰科植物铁皮石斛的显微结构,利用光镜和改进的蔗糖保护--液氮速冻冰冻切片法,观察铁皮石斛根、茎、叶的显微结构。该技术方法是将铁皮石斛器官经过蔗糖磷酸缓冲液保护液处理后抽真空,再经过液氮速冻、包埋、切片、展片观察、染色以及拍照等步骤,制作出铁皮石斛根、茎和叶较完整的显微结构切片。研究结果表明,适合铁皮石斛根的最适条件为:蔗糖质量体积分数为8%、冷凝温度-25℃、切片厚度20μm;茎的最适条件为:蔗糖质量体积分数为16%、冷凝温度-20℃、切片厚度15μm;叶的最适条件为:蔗糖质量体积分数为4%、冷凝温度-20℃、切片厚度10μm。该研究在兰科植物显微结构观察和组织化学研究中将具有广阔的应用前景。

In order to observe the microscopic structure of Dendrobium officinale quickly and efficiently. This study used the light microscope and the improved sucrose protection-liquid nitrogen quick-freezing freezing section method for orchid tissues and organs,Which improved sucrose protection-liquid nitrogen frozen method,observed root,stem and leaf of microscopic structure on D.officinale . The technology and methods treated D.officinale with sucrose phosphate buffer protection fluid,and then Vacuum processing. By liquid nitrogen frozen,embedding,sectioning,exhibition slice observation,staining and photo produced D.officinale roots,stems and leaves a complete microscopic section.The results show that the optimal condition for root was 8% sucrose mass volume fraction,condensation temperature -25℃,slice thickness 20 μm;the optimal condition for stem was 16% sucrose mass volume fraction,frozen temperature -20℃ and slice thickness 15 μm;the optimal condition for leaf was 4% sucrose mass volume fraction,frozen temperature -20℃ and slice thickness 10 μm. The study on the Orchidaceae microstructure observation and histochemical study will have broad prospect of application.