Marhavý Peter, Siddique Shahid
Umeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology, Swedish; University of Agricultural Sciences (SLU), Umeå, Sweden.
Department of Entomology and Nematology, University of California, Davis, One Shields Ave., Davis, CA, USA.
Bio Protoc. 2021 Feb 5;11(3):e3904. doi: 10.21769/BioProtoc.3904.
Histological stains are useful tools for characterizing cell shape, arrangement and the material they are made from. Stains can be used individually or simultaneously to mark different cell structures or polymers within the same cells, and to visualize them in different colors. Histological stains can be combined with genetically-encoded fluorescent proteins, which are useful for understanding of plant development. To visualize suberin lamellae by fluorescent microscopy, we improved a histological staining procedure with the dyes Fluorol Yellow 088 and aniline blue. In the complex plant organs such as roots, suberin lamellae are deposited deep within the root on the endodermal cell wall. Our procedure yields reliable and detailed images that can be used to determine the suberin pattern in root cells. The main advantage of this protocol is its efficiency, the detailed visualization of suberin localization it generates in the root, and the possibility of returning to the confocal images to analyze and re-evaluate data if necessary.
组织学染色是表征细胞形状、排列及其构成物质的有用工具。染色剂可单独使用或同时使用,以标记同一细胞内不同的细胞结构或聚合物,并以不同颜色使其可视化。组织学染色可与基因编码荧光蛋白相结合,这有助于了解植物发育。为了通过荧光显微镜观察木栓质片层,我们用荧光黄088和苯胺蓝染料改进了一种组织学染色程序。在根等复杂的植物器官中,木栓质片层沉积在根内皮层细胞壁的深处。我们的程序可产生可靠且详细的图像,可用于确定根细胞中的木栓质模式。该方案的主要优点在于其效率、在根中产生的木栓质定位的详细可视化,以及如有必要可返回共聚焦图像以分析和重新评估数据的可能性。