Phloem loading and long-distance transport of photoassimilate from source leaves to sink organs are essential physiological processes that contribute to plant growth and yield. At a minimum, three steps are involved: phloem loading in source organs, transport along the phloem path, and phloem unloading in sink organs. Each of these can have variable rates contingent on the physiological state of the plant, and thereby influence the overall transport rate. In addition to these phloem transport steps, rates of photosynthesis and photosynthate movement in the pre-phloem path, as well as photosynthate utilization in post phloem tissues of sink organs also contribute to phloem transport. The protocol described here estimates carbon allocation along the entire path from initial carbon fixation to delivery to sink organs after a labeling pulse: [14C]CO2 is photoassimilated in source leaves and loading and transport of the 14C label to heterotrophic sink organs (roots) is quantified by scintillation counting. This method is flexible and can be adapted to quantify long-distance transport in many plant species.