Objective: To describe online learning and eye strain situation of college students during the COVID-19 outbreak, to provide a scientific basis for guiding students eye health. Methods: A self-filled electronic questionnaire survey through questionnaire star was administered to college students across China. Information about online learning and eye strain of 1 046 college students during the epidemic was collected in Hefei, Anhui Province from March 16 to 20, 2020. The univariate and multivariate Logistic regression analysis were performed to analyze the association between online learning and eye strain of college students. Results: The rate of eye strain during online learning was 72.1%, totally of 68.4% in 421 male students and 74.6% in 625 female students. Boys with online learning time <6 h/d, slow internet access，difficulty in understanding online class reported higher rate of eye strain than girls( χ 2=17.36,8.72,7.02, P ＜0.05). Freshmen reported the highest rate of slow internet access occasionally and active online class( χ 2=15.26,16.11, P ＜0.05), junior students reported highest rate of online learning time <6 h/d, and easy understandable online class( χ 2=15.33,32.59, P ＜0.05), medical college students reported higher rate of slow internet access, inactive online class than non-medical college students( χ 2=11.79,11.03, P ＜0.05). Multivariate Logistic regression analysis showed that odds ratio( OR ) of eye strain in females was 1.40 (1.06-1.87), compared with males; the OR of eye strain were 1.43 (1.01-2.03) and 1.54 (1.10-2.15) in the groups with online learning time 6-<8 h/d and ≥8 h/d, respectively, compared with the group with online learning time <6 h/d, the OR of eye strain in the groups with slow internet access was 2.28 (1.25-4.14), compared with students without slow internet access, the OR of eye strain in the capable to understand and difficult to understand group were 2.54 (1.73-3.74) and 5.40 (2.70-10.80) respectively, compared with the easy to understand group. Conclusion Female students, online learing time ≥ 8 h/d, slow internet access, difficult to understand class content were positively related with college students eye strain. Attention should be paid to the eye health of college students to reduce the adverse effects of online learning on vision.during the COVID-19 epidemic.