AbstractAlthough manual restraint for small animal diagnostic radiography is common, worker protection is often not optimized, particularly for hands and eyes. Radiation safety training videos generally include hours of material on content other than personal protective equipment (PPE), have limited content, if any, on reducing dose to the lens of the eye, and are presented at the level of veterinary professionals. The objectives of this prospective, observational study were to develop a short, open access video training intervention at the layperson level, focused on proper use of PPE, and to test the effectiveness of the training video in changing behavior of workers. The use of PPE, optimal head position, and body position relative to the source of scattered radiation were compared before and after the video training was completed by workers. Results of unconditional and multivariable analyses were similar. In final multivariable analysis, workers wore gloves correctly more frequently (odds ratio [OR] = 2.09; 95% confidence interval [CI], 1.68‐2.59; P < .001) and wore eyewear more frequently (OR = 1.85; 95% CI, 1.23‐2.78; P = .003) after completing the training intervention. Workers also had an optimal head position more frequently (OR = 1.27; 95% CI, 1.03‐1.58; P = .03) and stood straight or leaned back more frequently (OR = 1.85; 95% CI, 1.48‐2.23; P < .001) after completing the training. The video training developed in this study is an effective tool that can be incorporated into a radiation protection program to improve worker radiation safety behaviors during manual restraint for small animal diagnostic radiography.