The use of learning technologies is becoming ubiquitous in higher education. As a result, there is a pressing need to develop methods to evaluate their accessibility to ensure that students do not encounter barriers to accessibility while engaging in e-learning. In this study, sample online units were evaluated for accessibility by automated tools and by student participants (in sessions moderated and unmoderated by researchers), and the data from these different methods of e-learning accessibility evaluation were compared. Nearly all students were observed encountering one or more barriers to accessibility while completing the online units, though the automated tools did not predict these barriers and instead predicted potential barriers that were not relevant to the study participants. These data underscore the need to carry out student-centered accessibility evaluation in addition to relying on automated tools and accessibility guideline conformance as measures of accessibility. Students preferred to participate in unmoderated sessions, and the data from the unmoderated sessions were comparable to that from the more traditional moderated sessions. Additional work is needed to further explore methods of student-centered evaluation, including different variations of unmoderated sessions.