利用整体染色与透明技术观察棉花胚囊方法

Using a whole stain-clearing technique to observe embryo sac of cotton

目的:探索一种观察棉花胚囊的方法。方法:用整体染色与透明技术对棉花胚囊进行处理,利用激光扫描共聚焦显微技术进行观察。首先在解剖镜下将内外珠被剥掉,取出胚囊;然后水解过程需要反复实验得到合适的时间,否则荧光染料很难进入细胞;最后材料在激光扫描共聚焦显微镜上用488nm激光激发观察并摄像。结果:可以清晰地观察到棉花胚囊组织的各组成部分及其细胞结构。结论:利用激光扫描共聚焦显微镜的重组功能构建胚囊的立体结构,有助于了解胚囊各组成细胞的空间结构,为观察棉花及其他较大子房或胚珠提供了一种简单快捷的方法。

Objective：To investigate a method using a whole stain-clearing technique for observations on embryo sac of cotton.Methods：With the whole stain-clearing technique to deal with cotton embryo sac and using confocal laser scanning microscopy observation.First,under anatomical lens inside and outside the bead will be peeled off,taking out the embryo sac,this step is critical,which needs a skilled process,otherwise,it is easy to both inside and outside,the stripping bead is broken for the embryo sac.Then,the hydrolysis process requires repeated experiments to get the right time,it is difficult to enter cells for fluorescent dye.Finally,the material on the laser scanning confocal microscope by using 488 nm laser excitation to observe and video.Results：It can be clearly observed to cotton embryo sac organization of each component and its cellular structure.Conclusion：It can build the three-dimensional structure of embryo sac,and contributes to an understanding of the spatial structure of embryo sac of each cell by using the restructuring of the confocal laser scanning microscopy.It provides a simple and fast method for observation of cotton and other large ovary or ovule.