水稻双受精过程的共聚焦显微镜观察

Observation on the double fertilization in rice by using confocal microscopy

(郑州大学离子束生物工程省重点实验室,郑州450052)摘要:首次利用核特异荧光染色与整体透明技术并利用激光扫描共聚焦显微镜(LSCM)对水稻双受精过程进行了观察。激光扫描共聚焦显微镜具有"组织与细胞CT"的功能,可以对整体组织进行扫描并构建三维结构。经荧光染色及透明处理后,在激光扫描共聚焦显微镜下经488nm激光激发,胚囊内各细胞的细胞核以及核仁发出明亮荧光,细胞核轮廓比较清晰,层次感强,并能清晰观察到胚囊内各细胞的结构特点和空间位置关系。不论是对开花前较小的成熟胚囊材料还是开花后较大的胚囊材料都取得了较好的观察效果。同传统的光学显微镜和荧光显微镜相比,其观察到的图像更清晰、更直观、更具立体感。

Nucleus fluorescent staining and whole clearing technique was first used in this study to observe the process of the double fertilization in rice by using Laser Scanning Confocal Microscopy （LSCM）. LSCM is a powerful tool in studying the structure of tissues and cells because of the function of＂CT Scanning＂. When the sample, which was stained with nucleus fluorescent and cleared with whole clearing technique,was excited by 488 nm laser,the nucleus and nucleolus of the cells in it excited bright fluorescent color and the profile of the nucleus was very clear. And the LSCM image showed perfectly the structure and special position of the cells in the embryo sac. Not only the images of small embryo sacs before fertilization, but also the images of big embryo sacs during and after fertilization were perfect. The results suggested that the LSCM was an ideal tool to study the process of double fertilization compared to traditional photics microscope.