Reproduction et Développement des Plantes, Université de Lyon, ENS de Lyon, CNRS, INRAE, Lyon Cedex 07, France.
Methods Mol Biol. 2022;2395:97-106. doi: 10.1007/978-1-0716-1816-5_6.
Growth and morphogenesis in plants depend on cell wall mechanics and on turgor pressure. Nanoindentation methods, such as atomic force microscopy (AFM), enable measurements of mechanical properties of a tissue at subcellular resolution, while confocal microscopy of tissues expressing fluorescent reporters indicates cell identity. Associating mechanical data with specific cells is essential to reveal the links between cell identity and cell mechanics. Here we describe an image analysis protocol that allows us to segment AFM scans containing information on tissue topography and/or mechanics, to stitch several scans in order to reconstitute an entire region of the tissue investigated, to segment the scans and label cells, and to associate labeled cells to the projection of confocal images. Thus all mechanical data can be mapped to the corresponding cells and to their identity. This protocol is implemented using NanoIndentation, a plugin that we are developing in the Fiji distribution of ImageJ.
植物的生长和形态发生依赖于细胞壁力学和膨压。纳米压痕方法，如原子力显微镜（AFM），可以在亚细胞分辨率下测量组织的力学性能，而表达荧光报告基因的组织的共焦显微镜则表明细胞的身份。将力学数据与特定细胞相关联对于揭示细胞身份和细胞力学之间的联系至关重要。在这里，我们描述了一种图像分析方案，该方案允许我们分割包含组织形貌和/或力学信息的 AFM 扫描，以便将几个扫描缝合在一起，以重新构建所研究组织的整个区域，分割扫描并标记细胞，并将标记的细胞与共焦图像的投影相关联。因此，所有的力学数据都可以映射到相应的细胞及其身份。该方案使用我们在 Fiji 版 ImageJ 中开发的 NanoIndentation 插件来实现。
