拟南芥花粉管的原位冷冻扫描电镜实验方法

花粉管在双受精过程中发挥着至关重要的作用,因此观察花粉管的生长是进行相关研究的重要手段.冷冻扫描电镜(Cryo-SEM)可实现对花粉管的原位观察,但由于花粉管样品的特殊性,常规制样方法并不适用.为此,研究者针对不同生长阶段的花粉管设计了两种冷冻扫描电镜的原位观察方法,并证实采用插秧式粘贴方法适用于柱突处的花粉管、采用液氮剥离方法适用于胎座表面处的花粉管,二种方法均可成功观察到花粉管的原位和原貌信息,为花粉管行为和相关受精机制的研究提供了有效方法.

In-situ experimental method on pollen tube of Arabidopsis by Cryo-SEM

Pollen tube plays criticle roles in double fertilization in angiosperms. The observation of pollen tube growth helps a lot in study on reproduction. Cryo-Scanning electron microscopy ( Cryo-SEM) can be applied to in-situ observation of pollen tube, but regular methods for sample preparation are not suitable. Accordingly, we designed two protocols by Cryo-SEM for pollen tubes in differernt developmental stages. The results showed that both transplanting-paste for pollen tube on stigma and stripping in liquid nitrogen for pollen tube on replum surface worked well to observe in-situ appearance and behavior of pollen tubes, which provided an effective pathway to study pollen tube growth and further double fertilization.