Mental fatigue measurement techniques utilize one or a combination of the cognitive, affective, and behavioral responses of the body. Eye-tracking and electrooculography, which are used to compute eye-based features, have gained momentum with increases in accuracy and robustness of the lightweight equipment emerging in the markets and can be used for objective and continuous assessment of mental fatigue. The main goal of this systematic review was to summarize the various eye-based features that have been used to measure mental fatigue and explore the relation of eye-based features to mental fatigue. The review process, following the preferred reporting items for systematic reviews and meta-analyses, used the electronic databases Web of Science, Scopus, ACM digital library, IEEE Xplore, and PubMed. Of the 1,385 retrieved documents, 34 studies met the inclusion criteria, resulting in 21 useful eye-based features. Categorizing these into eight groups revealed saccades as the most promising category, with saccade mean and peak velocity providing quick access to the cognitive states within 30 min of fatiguing activity. Complex brain networks involving sympathetic and parasympathetic nervous systems control the relation of mental fatigue to tonic pupil size and have the potential to indicate mental fatigue in controlled experimental conditions. Other categories, like blinks, are derived from the field of sleep research and should be used with caution. Several limitations emerged in the analysis, including varied experimental methods, use of dim lighting during the experiment (that could possibly also induce sleepiness), and use of unclear data analysis techniques, thereby complicating comparisons between studies.