It has been well established that the optimization of workflow can have substantial impact on the feasibility, efficiency, quality, safety, and outcomes of healthcare delivery. The process of studying and understanding workflow in support of such optimization has existed for some time, originally developed in the business and industrial research domains, and has been variably applied within the healthcare domain to date. In this chapter, we introduce the basic nomenclature and methods that encapsulate such workflow studies in the healthcare setting and provide a series of examples that demonstrate how such methods can be applied to solving critical problems. We conclude by reviewing open and active areas of inquiries concerning the current and future use of such workflow analysis methodologies.