Objective: The objective of this study was to analyze changes in the frequency and incidence of dry eye disease (DED) and screen exposure times during the onset of online courses, moreover, compare these changes between sexes. Secondarily, correlate online courses screen exposure times with the severity of DED symptoms. Methods: Four identical surveys, each containing the ocular surface disease index (OSDI) which quantifies DED symptom severity, and questions which categorized screen exposure times, were applied throughout 6 weeks. University students who had transitioned from face-to-face courses to online platforms were included in the study. Results: DED frequency among the 97 subjects (54 women and 43 men) peaked on week 4 (82.47%). OSDI scores significantly increased throughout the study (p < 0.0001) (baseline, 27.01 ± 17.55 versus Week 6, 37.17 ± 24.64), reflecting symptom worsening along an incidence of 8.5%. This worsening of symptoms occurred with women (p < 0.0001), while, in male subjects, it did not (p = 0.11); significant differences between sexes were found during the baseline (p = 0.01), Week 2 (p = 0.02), and Week 6 (p = 0.008), but not on Week 4 (p = 0.11). Online courses onset significantly increased screen exposure time (p < 0.0001). The baseline hours were 25.52 ± 11.33 and peaked on Week 2, being 34.62 ± 10.90. OSDI scores and online courses exposure times correlated significantly (Week 2, R = 0.265; Week 4, R = 0.262; and Week 6, R = 0.205). Conclusion: University students suffer from severe DED symptoms, which correlate with online courses onset. Educational institutions should foster ocular health.