This Guide supports state authorities in preventing and prosecuting radiological and nuclear (RN) crimes, which involve the illegal use of radioactive materials by organized crime groups and terrorists to cause mass panic and disruption. Despite stringent regulations, the accessibility of RN materials for malicious purposes poses a significant risk, exacerbated by the intersection of crime and terrorism, regional instabilities, and historical legacies of some states. The guide offers step-by-step advice on the entire process from investigation to court adjudication, recognizing the complexity and rarity of such cases. It includes practical tips, lessons learned, and best practices from real cases to help prosecutors build strong, evidence-based cases. A comprehensive training package developed by UNICRI includes modules, workshops, and mock trials to train prosecutors and judicial professionals, promoting the integration of these practices into national training curricula and enhancing global security.