The presence of biofilms in enclosed pipelines can lead to numerous deleterious issues. To date, it has been difficult to use optical imaging techniques to monitor the macroscale spatial distributions of biofilms. To address this concern, a combination of industrial computed tomography (ICT) and a contrast agent was explored to noninvasively visualize biofilms in three types of drip irrigation emitters. The results showed that ICT successfully observed and quantified the macroscale spatial distributions of biofilms. The complex hydrodynamic characteristics in the emitter channels affected the local distributions of biofilms. Biofilms were mainly attached to the lateral and medial faces and biomass decreased along the flow directions. Based on the distributions of biofilms, some emitter structural design defects were further diagnosed. Applying ICT in combination with the contrast agent could potentially provide a visual and effective way to reveal the formation mechanisms of biofilms and to optimize flow channel structures to avoid biofilm accumulations.