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A 3-D microwave imaging system prototype and an inverse scattering algorithm are developed to demonstrate the feasibility of 3-D microwave imaging for medical applications such as breast cancer detection with measured data. In this experimental prototype, the transmitting and receiving antennas are placed in a rectangular tub containing a fluid. The microwave scattering data are acquired by mechanically scanning a single transmit antenna and a single receive antenna, thus avoiding the mutual coupling that occurs when an array is used. Careful design and construction of the system has yielded accurate measurements of scattered fields so that even the weak scattered signals at S21 = -90 dB (or 30 dB below the background fields) can be measured accurately. Measurements are performed in the frequency domain at several discrete frequencies. The collected 3-D experimental data in fluid are processed by a 3-D nonlinear inverse scattering algorithm to unravel the complicated multiple scattering effects and produce high-resolution 3-D digital images of the dielectric constant and conductivity of the imaging domain. Dielectric objects as small as 5 mm in size have been imaged effectively at 1.74 GHz.