Radially configured microendoscopic electrical impedance probes intended for intraoperative surgical margin assessment during robot-assisted laparoscopic prostatectomy (RALP) were examined through simulation, bench-top experimentation, and ex vivo tissue studies. Three probe designs with 8, 9, and 17 electrodes, respectively, were analyzed through finite element method based simulations. One mm diameter spherical inclusions ( σinclusion = 1 S/m) are positioned at various locations within a hemispherical background ( σbackground = 0.1 S/m) of radius 5 mm. An 8-electrode configuration is not able to localize the inclusion at these positions while 9 and 17-electrode configurations are able to accurately reconstruct the inclusion at maximum depth of 1 mm and 3 mm, respectively. All three probe designs were constructed and evaluated using saline phantoms and ex vivo porcine and human prostate tissues. The 17-electrode probe performed best in saline phantom studies, accurately reconstructing high contrast, 1-mm-diameter metal cylindrical inclusions in a saline bath ( σsaline = 0.1 S/m) with a position and area error of 0.46 mm and 0.84 mm2, respectively. Additionally, the 17-electrode probe was able to adequately distinguish cancerous from benign tissues in three ex vivo human prostates. Simulations, bench-top saline experiments, and ex vivo tissue sampling suggest that for intraoperative surgical margin assessment during RALP, the 17-electrode probe (as compared to an 8 and 9 electrode probe) will be necessary to provide sufficient accuracy and sensitivity.