Faculty of Agriculture, Department of Agricultural Biotechnology, Eskisehir Osmangazi University, Eskisehir, Turkey.
Noble Research Institute, LLC, Ardmore, OK, USA.
Methods Mol Biol. 2022;2368:153-163. doi: 10.1007/978-1-0716-1677-2_11.
Microscopy is an important tool used for biological research and has played a crucial role toward understanding of cellular mechanisms and protein function. However, specific steps in processing of biological samples for microscopy warrant improvements to consistently generate data that can more reliably help in explaining mechanisms underlying complex biological phenomenon. Due to their small and fragile nature, some biological specimens such as Arabidopsis thaliana roots are vulnerable to damage during long sample preparation steps. Moreover, when specimens with a small diameter (typically less than 100 μm) are embedded in conventional silicone mold or capsule embedding, it is not only difficult to locate their orientation inside the capsule, but also a challenge to obtain good median longitudinal sections. Specimen orientation in particular is crucial because understanding certain plant biological processes such as gravitropism rely on precisely knowing spatial information of cells and tissues of the plant organ being studied. Here, we present a simple embedding technique to properly orient small plant organs such as roots so that the desired sectioning plane is achieved. This method is inexpensive and can be accomplished with minimal equipment and supplies.
显微镜是用于生物研究的重要工具，对于理解细胞机制和蛋白质功能发挥了关键作用。然而，为了进行显微镜检查而对生物样本进行的特定处理步骤需要改进，以持续生成更可靠的数据，从而更好地解释复杂生物现象背后的机制。由于其体积小且脆弱，一些生物标本（如拟南芥根）在长时间的样本准备过程中容易受损。此外，当直径较小（通常小于 100μm）的标本嵌入传统的硅胶模具或胶囊中时，不仅难以确定其在胶囊内的方向，而且很难获得良好的中纵切片。标本的方向尤其重要，因为要了解某些植物生物学过程，如向重力性，就必须精确了解正在研究的植物器官的细胞和组织的空间信息。在这里，我们提出了一种简单的包埋技术，可以正确定位小植物器官，如根，从而实现所需的切片平面。该方法成本低廉，只需最少的设备和耗材即可完成。
