A method to perform quantitative lesion analysis in molecular breast imaging (MBI) was developed using the opposing views from a novel dual-head dedicated gamma camera. Monte Carlo simulations and phantom models were used to simulate MBI images with known lesion parameters. A relationship between the full widths at 25%, 35%, and 50% of the maximum of intensity profiles through lesions and the true lesion diameter as a function of compressed breast thickness was developed in order to measure lesion diameter. Using knowledge of compressed breast thickness and the attenuation of gamma rays in soft tissue, a method was developed to measure the depth of the lesion to the collimator face. Using the measured lesion diameter and measurements of counts in the lesion and background breast region, relative radiotracer uptake or tumor to background ratio (T/B ratio) was calculated. Validation of the methods showed that the size, depth, and T/B ratio can be accurately measured for a range of small breast lesions with T/B ratios between 10:1 and 40:1 in breasts with compressed thicknesses between 4 and 10 cm. Future applications of this work include providing information about lesion location in patients for performing a biopsy of site and the development of a threshold for the T/B ratio that can distinguish benign from malignant disease.