The greatest challenge to guarding against radiological threats such as, radiological dispersal device (RDD) detonation or the use of a radiological emission device (RED), is the sheer prevalence and use of radioactive materials in facilities like academic institutions and medical centers. Securing these materials can be a daunting challenge. Cultivating and promoting a robust facility nuclear and radiological security culture, combined with other physical protection systems can assist in the prevention of malicious acts using radioactive materials. This promotion is highly dependent on a relevant self-assessment tool that would assess and correct deficiencies in organizational security culture. This paper investigates a series of culture indicators by assessing a wide range of medical centers and academic institutions across the United States. The study uses a quantitative method of online surveys to measure current perceptions and identify areas of strengths and weakness in particular aspects of security culture. Respondents to the survey include technicians, nurses, and other authorized users of radioactive materials at medical centers and radiation safety staff and students from academic institutions. The study attempts to examine the influence of human factors in the current state of emergency preparedness, security violations, preventive education and training, policy, and management oversight. The resulting outcome of the analysis outlines appropriate recommendations for facility-based security culture development, provides a process for raising security awareness, and promotes the sharing of best practices.