Online educational technologies and e-learning tools are providing new opportunities for students to learn worldwide, and they continue to play an important role in anatomical sciences education. Yet, as we shift to teaching online, particularly within the anatomical sciences, it has become apparent that e-learning tool success is based on more than just user satisfaction and preliminary learning outcomes-rather it is a multidimensional construct that should be addressed from an integrated perspective. The efficiency, effectiveness and satisfaction with which a user can navigate an e-learning tool is known as usability, and represents a construct which we propose can be used to quantitatively evaluate e-learning tool success. To assess the usability of an e-learning tool, usability testing should be employed during the design and development phases (i.e., prior to its release to users) as well as during its delivery (i.e., following its release to users). However, both the commercial educational software industry and individual academic developers in the anatomical sciences have overlooked the added value of additional usability testing. Reducing learner frustration and anxiety during e-learning tool use is essential in ensuring e-learning tool success, and will require a commitment on the part of the developers to engage in usability testing during all stages of an e-learning tool's life cycle. Anat Sci Educ 10: 190-199. © 2016 American Association of Anatomists.