ABSTRACT: Accurately assessing physical activity behavior in children, older adults, and adults with functional limitations is essential to further our understanding of determinants of physical activity behavior in these populations and to design, implement, and evaluate interventions designed to increase physical activity participation. Objective methods to assess physical activity behavior, owing to improvements in accuracy and precision over self-report measures, have become common in research and practice settings. This article reviews the current use of objective methods to assess physical activity in observational, determinant, and intervention studies for children, older adults, and adults with functional limitations. Important considerations are presented when adopting prediction algorithms developed on one population, and using in another population that is markedly different in age, health, and functional status. Best practices are presented, along with future recommendations for research to advance this area of scientific inquiry.