This paper describes the process and tools developed as part of a multidisciplinary collaborative simulation-based approach for iterative design and evaluation of operating room (OR) prototypes. Full-scale physical mock-ups of healthcare spaces offer an opportunity to actively communicate with and to engage multidisciplinary stakeholders in the design process. While mock-ups are increasingly being used in healthcare facility design projects, they are rarely evaluated in a manner to support active user feedback and engagement. Researchers and architecture students worked closely with clinicians and architects to develop OR design prototypes and engaged clinical end-users in simulated scenarios. An evaluation toolkit was developed to compare design prototypes. The mock-up evaluation helped the team make key decisions about room size, location of OR table, intra-room zoning, and doors location. Structured simulation based mock-up evaluations conducted in the design process can help stakeholders visualize their future workspace and provide active feedback.