To evaluate hygienic conditions using automatic injectors in magnetic resonance imaging (MRI) and computed tomography (CT) during clinical routine. The surfaces of medical devices (e. g., control console) and the palms of the technical and medical staff were microbiologically analyzed by taking imprints before and after hygienic education. In addition, the injector syringes for contrast medium (CM) and saline were checked for microbiological contamination following multiple (MRI: 14 h; CT 8 h) and single use. Furthermore, the potential of retrograde contamination from the patient along the tube was analyzed. A bacterial contamination with typical dermal bacteria was documented for the surfaces of the medical devices, the palms of the technical and medical staff, and the injection syringes following multiple use (MRI: 10/10 CM syringes, 6/10 saline syringes; CT: 8/10 CM syringes, 5/10 saline syringes). Correct hand disinfection in combination with single use of syringes avoided bacterial colonization. Retrograde bacterial contamination from the patient was not observed. Regular hygienic teaching sessions for technical and medical staff in MRI and CT departments using automatic injectors should be mandatory. Furthermore, the multiple use of syringes should be avoided until investigations addressing the potential of bacterial contamination are performed.