An optimization of free-hand sectioning of fine roots using fluorescence microscopy

Background and Aims High quality histological sections are crucial for observing plant structures. However, traditional procedures involving fixation, dehydration, embedding, clearing and staining are often laborious and time consuming, especially for the tiny fine root samples. Thus, simple, cheap and time-saving methods are necessary. Methods In this study, improved procedures were proposed for the quick and high-quality sections, including sectioning in soaked rushes (Juncus L.) stems and observation under a fluorescence microscope using DAPI filter, without staining and clearing. This modified method was applied to the fine root sectioning across six woody and five herbaceous species. Key Results This method allowed effective anatomical observation in both primary and secondary roots, especially for the xylem conduits. It is also worthy to note that the early sectioning practice is essential for the final anatomical observation. Conclusions This modified method free of chemical reagents and without clearing and staining, provided an optimization for good quality sections and rapid visualization of tiny root anatomy structures.