Aging is associated with a progressive decline in physical function characterized by decreased mobility, which is an important risk factor for loss of independence and reduced quality of life. Functional testing conducted in animals has advanced our understanding of age-related changes in physical ability and contributed to the development of physiologic measurements that can be used to assess functional changes during aging. The balance beam test is one assessment tool used to measure age-related changes in balance and coordination. The goal of this study is to provide analytical examples and psychometric support of a protocol that has been analyzed to show how the number of successive test runs, foot slips, pauses, and hesitations affect the reliability of the primary outcome measure, which is the time to cross the beam. Our results suggest that conducting more than 1 training session, consisting of greater than or equal to 3 successful training runs, followed by at least one test session with no less than 2 successful runs (that is, runs without pauses or hesitations) provides a psychometrically sound outcome. The data presented here indicate that a psychometric approach can improve protocol design and reliability of balance beam measures in mice.