Objective To investigate the effect of different flickering light frequencies for inducing myopia in C57BL/6 mice.Methods It was a animal experimental study.Ninety mice at age 28 days with similar refractions in both eyes were randomly assigned to five groups of 15 mice each.The control group was exposed to white non-flickering light.Three groups were exposed to white flickering lights at a high frequency,middle frequency,and low frequency at 10,5,and 2 Hz,respectively.A form-deprivation group consisted of 30 mice that underwent monocular form-deprivation with a semitransparent hemispherical thin plastic shell covering the right eyes,and was exposed to a non-flickering white light.Refraction and axial length were measured by murine-specific eccentric infrared photorefraction and A-scan ultrasonography at baseline and after two weeks of exposure.The data were analyzed by one way ANOVA and LSD t methods.Results ①The change in refraction and axial length in mice:the baselines of the refractions of the control group,high frequency,middle frequency and low frequency flickering light groups and the form-deprivation group were 2.627±0.494 D,2.597±0.626 D,2.552±0.558 D,2.617±0.568 D,2.590±0.657 D,respectively.The baselines of the axial length for the corresponding groups were 2.805±0.029 mm,2.803±0.026 mm,2.804±0.029 mm,2.806±0.026 mm,2.804±0.025 mm,respectively.There was no significant difference in refraction or axial length among the five groups (P＞0.05).After 2 weeks of exposure,the refraction of the control group,the high frequency,middle frequency,low frequency flickering light groups and the form-deprivation group were 3.720-±0.555 D,-1.710±0.632 D,-2.020±0.705 D,-2.612±0.686 D,-2.873±0.670 D,respectively.The axial lengths for the corresponding groups were 2.924±0.022 mm,2.995±0.024 mm,3.000±-0.020 mm,3.017±0.029 mm,3.026±0.016 mm,respectively.There was a significant difference in refraction and axial length between the control group and the other groups (P＜0.01).However,the induced changes in refraction and axial length were greater in the low frequency flickering light group and the form-deprivation group compared to the high and middle frequency flickering light groups.Conclusion B6 mice could be induced to form myopia by flickering light at all frequencies we used,and the low frequency of 2 Hz was the most effective in inducing myopia that is equivalent to form-deprivation myopia.