The factors and its mechanism which influence 4f imaging systemic resolution in practice were studied to guide the design and adjustment of the system. Firstly, the influence of the aberration caused by adjustment error on the system optical transfer function (MTF) in theory, whereby the mathematic model for describing the relationship of them was established. Based on the model, the adjustment error of 4f optical imaging system of 1 μm resolution was derived, and the influence of lens eccentricity, incline and displacement on the MTF was simulated, thus the variations of the resolution with the errors were achieved. The result indicates that within the incline of 15″, the eccentricity error is 0.02 mm and the displacement is within 0.01 mm. The adjustment error results in the decline of the resolution. Although the resolution increases while the incline within a minimum scope, the total trend will not change. The relationship figure of adjustment error and resolution can provide the reference for the error estimation of high-resolution 4f imaging system.