Vaccines are temperature-sensitive, and they may lose effectiveness without correct storage. Research in remote Australia raised questions regarding the integrity of the vaccine cold chain to Aboriginal communities and prompted the development of a video resource, Vaccine Story, to raise awareness of maintaining the cold chain. This study describes the development and evaluation of the Vaccine Story video. Local engagement helped to refine and produce the video. Ethics approval was gained from the Menzies School of Health Research Human Ethics Committee (20–3807) and the Central Australian Human Ethics Committee (17–2869) in 2020. The evaluation was considered low risk to all participants. An online anonymous survey tested the appropriateness and effectiveness of the Vaccine Story video to enhance knowledge and awareness of risks to the vaccine cold chain and the potential implementation of the Vaccine Story video. Diverse health professionals and other staff (n = 83) completed the survey. Responses highlighted a lack of training around the packing and transport of vaccines, particularly for non-clinical support staff. Respondents stated the health messaging in the Vaccine Story video was effective. Over half of respondents (52%) recommend the use of the video as a training tool. The Vaccine Story video addresses some concerns regarding vaccine transportation and storage in remote Aboriginal communities through strengthening understanding of the importance of cold chain.