We have developed a breast tomosynthesis system utilizing a selenium-based direct conversion flat panel detector. This prototype system is a modification of Selenia, Hologic’s full field digital mammography system, using an add-on breast holding device to allow 3D tomosynthetic imaging. During a tomosynthesis scan, the breast is held stationary while the x-ray source and detector mounted on a c-arm rotate continuously around the breast over an angular range up to 30 degrees. The x-ray tube is pulsed to acquire 11 projections at desired c-arm angles. Images are reconstructed in planes parallel to the breastplate using a filtered backprojection algorithm. Processing time is typically 1 minute for a 50 mm thick breast at 0.1 mm in-plane pixel size, 1 mm slice-to-slice separation. Clinical studies are in progress. Performance evaluations were carried out at the system and the subsystem levels including spatial resolution, signal-to-noise ratio, spectra optimization, imaging technique, and phantom and patient studies. Experimental results show that we have successfully built a tomosynthesis system with images showing less structure noise and revealing 3D information compared with the conventional mammogram. We introduce, for the first time, the definition of “Depth of Field” for tomosynthesis based on a spatial resolution study. This parameter is used together with Modulation Transfer Function (MTF) to evaluate 3D resolution of a tomosynthesis system as a function of system design, imaging technique, and reconstruction algorithm. Findings from the on-going clinical studies will help the design of the next generation tomosynthesis system offering improved performance.