An Ussing chamber was designed for impedance analysis of epithelial tissue and optimized for the use of high-frequency alternating current stimuli. Shielded voltage electrodes, located axially within the electric field of the Ussing chamber, minimized the reactive properties of the set-up. By vectorial subtraction, the small reactive contribution of the optimized Ussing chamber was completely compensated for. This method allowed for transmural impedance measurements in a frequency range of 1-65 kHz. For the first time, Nyquist plots of cultured intestinal cell monolayers (HT-29/B6) are presented. The epithelial monolayers in different stages of confluence showed the impedance locus as a semicircle, with the high frequency end close to the origin. These epithelial monolayers could be modeled by a simple RC-parallel circuit without a series resistance.