Ullah, M. N.; Hastings, D.; Lee, S. J.; Park, W.; Zou, S. J.; Anders, D.; Park, J. H.; Weisenberger, D.; Cheng, W.; Abbaszadeh, S.; Levin, C.

Imaging carbon movements in the rhizosphere is fundamentally limited by high soil heterogeneity, low signal levels, and lack of methodology. We present Rhizo-PET, a dedicated positron emission tomography (PET) imaging and analysis framework designed to characterize the 4D spatiotemporal patterns of tracer distribution in intact plant-soil systems. The system achieved a global energy resolution of 11.93 {+/-} 0.02% FWHM at 511 keV and maintained stable performance over 8 h of continuous acquisition, with a coincidence rate variation of only 0.7%. Spatial resolution reached 1.06 mm near the center of the field of view, establishing a high-fidelity region for root-scale analysis. Dynamic datasets were acquired from live Phaseolus vulgaris plants (N = 3) over 180 min following 11CO2 pulse labeling and reconstructed into 3 min temporal frames. Quantitative analysis across 243 independent regions of interest (ROI) revealed that cumulative tracer accumulation decreases monotonically with radial distance from the root axis, while axial transport delays increase systematically in lower root segments (p < 0.001). Hierarchical variability analysis showed that within-plant spatial organization (CVTTP = 0.03) is significantly more stable than inter-plant variation (CVTTP = 0.14), proving that the observed heterogeneity reflects biological spatial organization rather than experimental instability. These results establish Rhizo-PET as a robust, reproducible platform for the non-invasive, time-resolved analysis of carbon dynamics in the rhizosphere under realistic soil conditions.