Diffuse optical spectroscopy (DOS) and imaging are emerging diagnostic techniques that quantitatively measure the concentration of deoxy-hemoglobin (ctHHb), oxy-hemoglobin (ctO 2Hb), water (ctH 2O), and lipid in cm-thick tissues. In early-stage clinical studies, diffuse optical imaging and DOS have been used to characterize breast tumor biochemical composition and monitor therapeutic response in stage II/III neoadjuvant chemotherapy patients. We investigated whether DOS measurements obtained before and 1 week into a 3-month adriamycin/cytoxan neoadjuvant chemotherapy regimen can predict final, postsurgical pathological response. Baseline DOS measurements of 11 patients before therapy revealed significant increases in tumor ctHHb, ctO 2Hb, ctH 2O, and spectral scattering slope, and decreases in bulk lipids, relative to normal breast tissue. Tumor concentrations of ctHHb, ctO 2Hb, and ctH2O dropped 27 ؎ 15%, 33 ؎ 7%, and 11 ؎ 15%, respectively, within 1 week (6.5 ؎ 1.4 days) of the first treatment for pathology-confirmed responders (n ؍ 6), whereas nonresponders (n ؍ 5) and normal side controls showed no significant changes in these parameters. The best single predictor of therapeutic response 1 week posttreatment was ctHHb (83% sensitivity, 100% specificity), while discrimination analysis based on combined ctHHb and ctH 2O changes classified responders vs. nonresponders with 100% sensitivity and specificity. In addition, the pretreatment tumor-to-normal ctO 2Hb ratio was significantly higher in responders (2.82 ؎ 0.44) vs. nonresponders (1.82 ؎ 0.49). These results highlight DOS sensitivity to tumor cellular metabolism and biochemical composition and demonstrate its potential for predicting and monitoring an individual's response to treatment.diffuse optical imaging ͉ frequency-domain photon migration ͉ near-infrared ͉ tissue spectroscopy ͉ translational research O ptimal management locally advanced breast cancer (LABC) remains a complex therapeutic problem (1). LABC represents 5-20% of all newly diagnosed breast cancers in the United States with a higher incidence in medically underserved areas (2). Treatment for LABC has evolved from radical mastectomy to preoperative neoadjuvant chemotherapy followed by mastectomy or breast conservation therapy (3). Despite aggressive local therapy, long-term patient survival is still poor. LABC remains controversial because of uncertainties in determining the optimal intensity and duration of neoadjuvant chemotherapy and evaluating therapeutic response (2, 4, 5).Neoadjuvant chemotherapy response is determined by serial physical examination, mammography and/or ultrasound. Complete pathological response (cPR) is an important therapeutic endpoint that is a surrogate for eradicating micrometastases, and strongly correlates with patient survival (6). Thus one goal of neoadjuvant chemotherapy monitoring is to determine early when a patient will demonstrate cPR. Many studies revealed significant discrepancies between clinical response assessments and final pathology (7-9).A rece...