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Only recently has physical space design become more widely recognized as playing a critical role in delivery of care, with an emerging body of literature on the application of human factors approaches to design and evaluation. This chapter describes the use of human factors approaches to develop and conduct an evaluation of a proposed Neonatal Intensive Care Unit redesign in a Midwestern children’s hospital. Methods included observations and knowledge elicitation from stakeholders to characterize their goals, challenges, and needs. This characterization is integral to informing the design of user-centered solutions, including physical space design. We also describe an approach to evaluating the proposed design that yielded actionable recommendations specific to hospital-driven design goals.