【作者】 宁代锋； 尹增芳； 张菁； 孙海燕；

【Author】 Ning Daifeng Yin Zengfang~* Zhang Jing Sun Haiyan(College of Forest Resources and Environment Nanjing Forestry University,Nanjing 210037)

【机构】 南京林业大学森林资源与环境学院；

【摘要】 利用常规电镜技术观察了冰冻切片过程中不同冷冻方法对植物细胞超微结构影响,研究发现利用直接包埋法处理的植物细胞超微结构保存较好,而液氮冷冻处理后植物细胞内膜系统损伤严重。因此,本文建立了一种直接包埋冷冻和适当回温相结合的方法,不仅可以制作出完整的植物组织切片,而且有助于植物细胞基本结构的保存。其操作程序是：固定与包埋→直接冷冻→适当回温→快速切片→展片。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。更多还原

【Abstract】 The ultrastructure of plant cells under different freezing treatments during cryosectioning were observed by using the transmission electron microscopy.It was found that the cell ultrastructure were well preserved with direct cryoembed,but the endomembrane system in the cell was injured severely with liquid nitrogen cryopreservation.So we established a method with combination of cryoembed and modest thawing,by which it not only can produce integrated plant tissue sections,but also preserve the basic structure of plant cells. The procedure consisted of several steps as follows:fixing and embeding→direct refrigeration→modest thawing→rapid sectioning→section flatting.The section made by direct cryoembed with modest thawing could keep on staihing and cell chemical determination in different ways.Besides,the characteristic of this procedure is simple and easy to be promoted.更多还原