Objective To explore the clinical application of molybdenum target non-stereoscopic guidewire guided excision in the removal of breast microcalcification foci and guiding surgical biopsy. Methods The patients with microcalcification of the breast were randomly divided into two groups according to the random number table. The two groups were manually positioned, and the lines were taken separately. All the specimens were taken for mammography, and the crochet and the nearest calcification point were measured. The distance is statistically analyzed. Results On average, each patient in the experimental group underwent mammography for 5.5 times. While in the control group, the mammography was 6.12 times per patient, which was statistically significant compared with the experimental group (P < 0.05). The distance between the crochet and the nearest calcification point is (6.13 ± 0.59) mm in the experimental group, and the distance between the crochet and the nearest calcification point was (6.79 ± 0.58) mm in the control group, which mean statistically significant between two groups (P < 0.05). No complications such as hemorrhage and localization of guide wire were found. The positioning process was smooth and one-time biopsy was successful during the operation. Conclusion The mammography Xray manual localization of mammary microcalcifi-cations is simple, easy, accurate, and has a satisfactory positioning effect.