Abstract Study objective The purpose of this study is to evaluate the usability of emergency department (ED) software prototypes developed for Tablet personal computers (Tablet PCs) in order to keep electronic health records (EHRs) of patients errorless and accessible through mobile technologies. In order to serve this purpose, two alternative prototypes were developed for Tablet PCs: Mobile Emergency Department Software (MEDS) and Mobile Emergency Department Software Iconic (MEDSI) among which the user might choose the more appropriate one for ED operations based on a usability analysis involving the target users. Methods The study is based on a case study of 32 potential users of our prototypes at the ED of Kadikoy-AHG in Istanbul, Turkey. We examined usability of the prototypes for medical information systems by means of Nielsen’s heuristic evaluation and cognitive walkthrough methods relying on 7-point scales, and scenario completion success rate and average scenario completion time, respectively. Results The implementation of MEDSI in our case study confirmed the view that the usability evaluation results of iconic GUIs were better than those of non-iconic GUIs in terms of Nielsen’s heuristic evaluation, effectiveness and user satisfaction. For the whole sample, paired t-test scores indicated that there was a significant difference ( p < 0.01) between mean values of Nielsen’s usability scores toward MEDS and MEDSI indicating that MEDSI was evaluated more favorably than MEDS. As for effectiveness of the prototypes, significant differences ( p < 0.01) were noted between MEDS and MEDSI in terms of both overall scenario completion success rate and average scenario completion time. Similarly, for the full sample of users independent sample t-test scores indicated that MEDSI was perceived significantly more favorable ( p < 0.01) than MEDS in terms of overall user satisfaction. Conclusion The study provides two important contributions to the extant literature. First, it addresses a topic and methodology that serves potentially interesting to the biomedical informatics community. Drawing on good background information and appropriate context, it involves various aspects of usability testing. Another contribution of the study lies in its examination of two different prototypes during the design phase involving the target users.