The purpose of this study was to evaluate the potential for use of image volume based registration (IVBaR) to aid in measurement of changes in the tumor during chemotherapy of breast cancer. Successful IVBaR could aid in the detection of such changes in response to neoadjuvant chemotherapy and potentially be useful for routine breast cancer screening and diagnosis. IVBaR was employed in a new method of automated estimation of tumor volume in studies following the radiologist identification of the tumor region in the prechemotherapy scan. The authors have also introduced a new semiautomated method for validation of registration based on Doppler ultrasound (U.S.) signals that are independent of the grayscale signals used for registration. This Institutional Review Board approved study was conducted on 10 patients undergoing chemotherapy and 14 patients with a suspicious/unknown mass scheduled to undergo biopsy. Reasonably reproducible mammographic positioning and nearly whole breast U.S. imaging were achieved. The image volume was registered offline with a mutual information cost function and global interpolation based on a thin‐plate spline using MIAMI FUSE© software developed at the University of Michigan. The success and accuracy of registration of the three dimensional (3D) U.S. image volume were measured by means of mean registration error (MRE). IVBaR was successful with MRE of in 9 out of 10 reproducibility automated breast ultrasound (ABU) studies and in 12 out of 17 ABU image pairs collected before, during, or after of chemotherapy. Semiautomated tumor volume estimation was performed on registered image volumes giving mean accuracy compared to the radiologist hand‐segmented tumor volume on seven cases. Doppler studies yielded fractional volume of color pixels in the region surrounding the lesion and its change with changing breast compression. The Doppler study of patients with detectable blood flow included five patients with suspicious masses and three undergoing chemotherapy. Spatial alignment of the 3D blood vessel data from the Doppler studies provided independent measures for the validation of registration. In 15 Doppler image volume pairs scanned with differing breast compression, the mean centerline separation value was , while MRE based on a few identifiable structural points common to the two grayscale image volumes was . Another measure, the overlap ratio of blood vessels, was shown to increase from 0.32 to 0.59 with IVBaR for pairs at various compression levels. These results show that successful registration of ABU scans may be accomplished for comparison and integration of information.