To achieve the consistency with subjective evaluation, the paper proposes a microscopy image sharpness metric using visual attention mechanism and edge spreading measurement. According to the gray difference of pixels at different spatial location, visual attention index map is calculated to simulate the way of human vision system observing the features of scenery. By this way, weight is given for different regions and pixels, and the potential focuing areas is extract by segmentation. Using edge spreading evaluation in this areas, an average edge spreading of horizontal and vertical direction is obtained, which is considered as the last sharpness metric. The authors design experiments for subjective assessment and objective evaluation. The experiment and comparison demonstrate that, the proposed method can effectively solve the key problems in the evaluation of microscopy image, and satisfy the consistency of objective and subjective evaluation.