Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), Versailles, France.
Methods Mol Biol. 2024;2830:93-104. doi: 10.1007/978-1-0716-3965-8_9.
In flowering plants, proper seed development is achieved through the constant interplay of fertilization products, embryo and endosperm, and maternal tissues. Understanding such a complex biological process requires microscopy techniques able to unveil the seed internal morphological structure. Seed thickness and relatively low permeability make conventional tissue staining techniques impractical unless combined with time-consuming dissecting methods. Here, we describe two techniques to imaging the three-dimensional structure of Arabidopsis seeds by confocal laser scanning microscopy. Both procedures, while differing in their time of execution and resolution, are based on cell wall staining of seed tissues with fluorescent dyes.
在开花植物中,通过受精产物、胚胎和胚乳以及母体组织的不断相互作用,实现了种子的正常发育。要理解这样一个复杂的生物学过程,就需要用到能够揭示种子内部形态结构的显微镜技术。种子的厚度和相对较低的渗透性使得传统的组织染色技术变得不切实际,除非结合费时的解剖方法。在这里,我们描述了两种通过共聚焦激光扫描显微镜对拟南芥种子的三维结构进行成像的技术。这两种方法虽然在执行时间和分辨率上有所不同,但都基于用荧光染料对种子组织进行细胞壁染色。