Abstract. We present a study of the bilateral symmetry of human breast hemoglobin saturation maps measured with a broadband optical mammography instrument. We have imaged 21 patients with unilateral breast cancer, 32 patients with unilateral benign lesions, and 27 healthy patients. An image registration process was applied to the bilateral hemoglobin saturation (SO 2 ) images by assigning each pixel to the low, middle, or high range of SO 2 values, where the thresholds for the categories were the 15th and 85th percentiles of the individual saturation range. The Dice coefficient, which is a measure of similarity, was calculated for each patient's pair of right and left breast SO 2 images. The invasive cancer patients were found to have an average Dice coefficient value of 0.55 AE 0.07, which was significantly lower than the benign and healthy groups (0.61 AE 0.11 and 0.62 AE 0.12, respectively). Although differences were seen in a group analysis, the healthy patient Dice coefficients spanned a wide range, limiting the diagnostic capabilities of this SO 2 symmetry analysis on an individual basis. Our results suggest that for assessing the SO 2 contrast of breast lesions, it may be better to select a reference tissue in the ipsilateral rather than the contralateral breast. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.