X-ray imaging of leaf venation networks.

Leaf venation networks mediate many plant resource fluxes and are therefore of broad interest to research questions in plant physiology, systematics, paleoecology, and physics. However, the study of these networks is limited by slow and destructive imaging methods. X-ray imaging of leaf veins is potentially rapid, of high resolution, and nondestructive. Here, we have developed theory for absorption- and phase-contrast X-ray imaging. We then experimentally test these approaches using a synchrotron light source and two commercially available X-ray instruments. Using synchrotron light, we found that major veins could be consistently visualized using absorption-contrast imaging with X-ray energies < 10 keV, while both major and minor veins could be consistently visualized with the use of an iodine contrast agent at an X-ray energy of 33.269 keV. Phase-contrast imaging at a range of energies provided high resolution but highlighted individual cell walls more than veins. Both approaches allowed several hundred samples to be processed per d. Commercial X-ray instruments were able to resolve major veins and some minor veins using absorption contrast. These results show that both commercial and synchrotron X-ray imaging can be successfully applied to leaf venation networks, facilitating research in multiple fields.