Non-destructive diffraction enhanced imaging of seeds.

Techniques that make possible the non-destructive continuous observation of plant anatomy and developmental processes provide novel insights into these phenomena. Non-destructive imaging of seeds was demonstrated using the synchrotron-based X-ray imaging technique, diffraction enhanced imaging (DEI). The seed images obtained had good contrast and definition, allowing anatomical structures and physiological events to be observed. Structures such as hypocotyl-root axes, cotyledons, seed coats, air cavities, and embryo-less Brassica napus L. seeds were readily observed using DEI. Embryo axes, scutella, pericarp furrows, coleoptiles, and roots were observable over a time-course in individual germinating Triticum aestivum L. caryopses. Novel anatomical and physiological observations were also made that would have been difficult to make continuously using other techniques. The physical principles behind DEI make it a unique imaging technique. Contrast in DEI is the result of X-ray refraction at the density differences occurring at tissue boundaries, scatter caused by regions containing ordered molecules such as cellulose fibres, and attenuation. Sectioning of samples and the infusion of stains or other contrast agents are not necessary. Furthermore, as high-energy X-rays are used (30-40 keV), little X-ray absorption occurs, resulting in low levels of radiation damage. Consequently, studies of developmental processes may be performed on individuals. Individual germinating B. napus and T. aestivum seeds were imaged at several time points without incurring any apparent radiation damage. DEI offers a unique way of examining plant anatomy, development, and physiology, and provides images that are complementary to those obtained through other techniques.