Abstract：
Arabidopsis thaliana developing pollen grains serve as an excellent system for studying vacuole dynamics. Here, we present a methodological approach that utilizes the serial tomography package in Etomo software from IMOD to generate whole-cell tomograms on A. thaliana developing pollens for visualizing vacuoles on the whole-cell scale. In order to understand the vacuole dynamics along with the pollen maturation, we also introduce a sampling method aimed at harvesting the pollen grains at various stages, marked by the vegetative nucleus or generative cell. The cryo-fixation/freeze-substitution technique can then be applied to preserve the fine structures of the pollen grains and facilitate detailed ultrastructure examination. Through this method, large-volume whole-cell electron tomograms regarding vacuolar morphologies and ultrastructural changes during pollen development and maturation have been obtained. Overall, the method presented here provides valuable insights into the dynamic nature of vacuoles in Arabidopsis developing pollen.
摘要：
拟南芥发育的花粉粒是研究液泡动力学的极好系统。这里,我们提出了一种方法学方法，该方法利用IMOD的Etomo软件中的串行断层扫描软件包在拟南芥上生成全细胞断层照片，以开发花粉，以在全细胞尺度上可视化液泡。为了了解随着花粉成熟的液泡动力学，我们还介绍了一种采样方法，旨在在各个阶段收获花粉粒，以营养核或生殖细胞为标志。然后可以应用冷冻固定/冷冻替代技术来保留花粉粒的精细结构并促进详细的超微结构检查。通过这种方法,已获得有关花粉发育和成熟过程中液泡形态和超微结构变化的大容量全细胞电子断层扫描图。总的来说,这里介绍的方法为拟南芥发育花粉中液泡的动态性质提供了有价值的见解。
