The purification and sorting of cells using microfluidic methodologies has been a remarkably active area of research over the past decade. Much of the scientific and technological work associated with microfluidic cell separation has been driven by needs in clinical diagnostics and therapeutic monitoring, most notably in the context of circulating tumor cells. The last several years have seen advances in a broad range of separation modalities ranging from miniaturized analogs of established techniques such as fluorescence- and magnetic-activated cell sorting (FACS and MACS, respectively), to more specialized approaches based on affinity, dielectrophoretic mobility, and inertial properties of cells. With several of these technologies nearing commercialization, there is a sense that the field of microfluidic cell separation has achieved a high level of maturity over an unusually short span of time. In this Perspective, we set the stage by describing major scientific and technological advances in this field and ask what the future holds. While many scientific questions remain unanswered and new compelling questions will undoubtedly arise, the relative maturity of this field poses some unique challenges.