In this chapter, we review scientific findings that form the basis for neuroimaging and neurophysiological biomarkers for ADHD diagnosis and treatment. We then highlight the different challenges in translating mechanistic findings into biomarkers for ADHD diagnosis and treatment. Population heterogeneity is a primary barrier for identifying biomarkers of ADHD diagnosis, which requires shifts toward dimensional approaches that identify clinically useful subgroups or prospective biomarkers that can identify trajectories of illness, function, or treatment response. Methodological limitations, including emphasis on group level analyses of treatment effects in small sample sizes, are the primary barriers to biomarker discovery in ADHD treatment. Modifications to clinical trials, including shifting towards testing biomarkers of a priori prediction of functionally related brain targets, treatment response, and side effects, are suggested. Finally, future directions for biomarker work are discussed.