Jones Dylan H, Atkinson Brian S, Ware Alexander, Sturrock Craig J, Bishopp Anthony, Wells Darren M
Integrated Phenomics Group, School of Biosciences, University of Nottingham, Nottingham, United Kingdom.
Hounsfield Facility, School of Biosciences, University of Nottingham, Nottingham, United Kingdom.
Front Plant Sci. 2021 Jan 8;11:617830. doi: 10.3389/fpls.2020.617830. eCollection 2020.
Quantification of anatomical and compositional features underpins both fundamental and applied studies of plant structure and function. Relatively few non-invasive techniques are available for aquatic plants. Traditional methods such as sectioning are low-throughput and provide 2-dimensional information. X-ray Computed Microtomography (μCT) offers a non-destructive method of three dimensional (3D) imaging , but has not been widely used for aquatic species, due to the difficulties in sample preparation and handling. We present a novel sample handling protocol for aquatic plant material developed for μCT imaging, using duckweed plants and turions as exemplars, and compare the method against existing approaches. This technique allows for previously unseen 3D volume analysis of gaseous filled spaces, cell material, and sub-cellular features. The described embedding method, utilizing petrolatum gel for sample mounting, was shown to preserve sample quality during scanning, and to display sufficiently different X-ray attenuation to the plant material to be easily differentiated by image analysis pipelines. We present this technique as an improved method for anatomical structural analysis that provides novel cellular and developmental information.
对植物结构和功能的基础研究与应用研究而言，解剖学和成分特征的量化至关重要。用于水生植物的非侵入性技术相对较少。诸如切片等传统方法通量较低，且只能提供二维信息。X射线计算机显微断层扫描（μCT）提供了一种三维（3D）成像的无损方法，但由于样本制备和处理存在困难，尚未广泛应用于水生物种。我们展示了一种专为μCT成像开发的用于水生植物材料的新型样本处理方案，以浮萍植物和块茎为示例，并将该方法与现有方法进行比较。这项技术能够对充满气体的空间、细胞物质和亚细胞特征进行前所未有的三维体积分析。所描述的嵌入方法，即利用凡士林凝胶进行样本固定，在扫描过程中能够保持样本质量，并且与植物材料显示出足够不同的X射线衰减，以便通过图像分析流程轻松区分。我们将这项技术作为一种改进的解剖结构分析方法呈现出来，它能提供新的细胞和发育信息。
