An optimization of freehand sectioning of fine roots using fluorescence microscopy

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.

In this study, improved procedures were proposed for quick and high-quality sections, including sectioning within the stems of soaked rushes (Juncus L.) and observation under a fluorescence microscope using a 4',6-diamidino-2-phenylindole filter, without staining and clearing. This modified method was applied to section fine roots of six woody and five herbaceous species.

This method allowed effective anatomical observation in both primary and secondary roots, especially for the xylem conduits. It is also noteworthy that early sectioning practice is essential for the final anatomical observation.

This modified method, sectioning within the stems of soaked rushes, free of chemical reagents and without clearing and staining, provided an optimization for good-quality sections and rapid visualization of tiny root anatomical structures.