Hedhly Afif, Vogler Hannes, Eichenberger Christof, Grossniklaus Ueli
Department of Plant and Microbial Biology, Zurich-Basel Plant Science Center, University of Zurich;
Department of Plant and Microbial Biology, Zurich-Basel Plant Science Center, University of Zurich.
J Vis Exp. 2018 Apr 12(134):56441. doi: 10.3791/56441.
Due to its formidable tools for molecular genetic studies, Arabidopsis thaliana is one of the most prominent model species in plant biology and, especially, in plant reproductive biology. However, plant morphological, anatomical, and ultrastructural analyses traditionally involve time-consuming embedding and sectioning procedures for bright field, scanning, and electron microscopy. Recent progress in confocal fluorescence microscopy, state-of-the-art 3-D computer-aided microscopic analyses, and the continuous refinement of molecular techniques to be used on minimally processed whole-mount specimens, has led to an increased demand for developing efficient and minimal sample processing techniques. In this protocol, we describe techniques for properly dissecting Arabidopsis flowers and siliques, basic clearing techniques, and some staining procedures for whole-mount observations of reproductive structures.
由于其在分子遗传学研究方面拥有强大工具，拟南芥是植物生物学尤其是植物生殖生物学中最突出的模式物种之一。然而，传统的植物形态学、解剖学和超微结构分析涉及用于明场、扫描和电子显微镜的耗时包埋和切片程序。共聚焦荧光显微镜、先进的三维计算机辅助显微镜分析的最新进展，以及用于最少处理的整装标本的分子技术的不断完善，导致对开发高效且最少样本处理技术的需求增加。在本方案中，我们描述了正确解剖拟南芥花和角果的技术、基本的透明技术，以及一些用于生殖结构整装观察的染色程序。
