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Non-invasive 3D methods for imaging cementum increments using synchrotron radiation sources are one of the most promising new avenues for cementum research. This technique offers the opportunity to overcome the major caveats to traditional thin section imaging, and provides volumetric datasets of sub-micrometer resolution that can be investigated in new ways. Such studies can unlock the 3D structure of cementum increments, and 3D measures may allow for new inferences on the relationship between cementum growth and life history. However, as a new field of research, synchrotron X-ray imaging of cementum must ensure reproducibility by employing quantitative approaches to develop optimal experimental procedures and settings for imaging cementum in different samples. The quantitative parameter optimisation procedure we introduce in this chapter should form a crucial part of the imaging protocol that we present here, in which we outline the major steps in preparing for, performing and concluding a synchrotron imaging experiment, based on our own experience.