Neutron Radiography (NR) is a valuable non-invasive tool to study in situ root development in soil. However, there is a lacuna of quantitative information on its capabilities and limitations. We combined neutron radiography with image analysis techniques to quantify the neutron absorption coefficients (Σ) of various root-growth media for a range of water contents (θ) in the presence and absence of plant roots with various rooting systems. Plants were grown in aluminium containers (170 × 150 × 12 mm) and were imaged using NR, as well as X-Ray radiography and an optical scanner. Sandy soil was the best medium for NR because it supported plant growth at θ that gave a good contrast for root visualisation. After correction for neutron scattering, we obtained a linear correlation between Σ and soil θ. The minimum detectable root thickness in neutron radiographs was found to be 0.2 mm in these containers. Combining NR with X-Ray radiography could provide information on soil structure in addition to revealing root structure and development.