Background: Despite increasing knowledge about the potential benefits of advanced user-controlled technology, the decision about switching an individual prosthesis user from a non-microprocessor prosthetic knee to a microprocessor prosthetic knee is mainly based on clinician’s experience rather than empirical evidence. Objectives: To demonstrate the utility of single-subject design and data analysis for evaluating changes in temporal-spatial gait characteristics between walking with a non-microprocessor prosthetic knee and microprocessor prosthetic knee. Study design: Single-subject ABA/BAB design. Methods: Seven non-microprocessor prosthetic knee users (all men, age 50–84 years, 3–40 years post-amputation) were transitioned through the ABA or BAB phases (A-NMPK, B-MPK, 5 weeks each). Four weekly gait evaluations were performed at three self-selected speeds with an electronic walkway. The non-microprocessor prosthetic knee–microprocessor prosthetic knee differences in stride length–cadence relationship, prosthetic weight acceptance, single-limb support, and step width were evaluated for each subject using the “non-overlap of all pairs” statistical method. Results: Most subjects improved temporal-spatial gait while on the microprocessor prosthetic knee; in only one subject, none of the 10 gait parameters were in favor of the microprocessor prosthetic knee. In the BAB group, longer use of the microprocessor prosthetic knee was associated with shorter prosthetic weight acceptance and longer single-limb support times across three speeds. Step width either improved with the microprocessor prosthetic knee or remained unchanged in most subjects. Conclusion: The evidence of individual subject improvements in gait coordination, greater reliance on the prosthetic side, and better stability with the microprocessor prosthetic knee than non-microprocessor prosthetic knee over a range of walking speeds demonstrate the practical utility of the single-subject method in clinical decision-making. Clinical relevance The results demonstrate the use of the single-subject method for examining person-specific differences in temporal-spatial gait characteristics between walking with a non-microprocessor prosthetic knee and microprocessor prosthetic knee at three self-selected speeds. The method proved feasible and reliable for documenting changes in gait at the individual level, which is relevant for clinical practice.