MFEIT (multi-frequency electrical impedance tomography) could distinguish between ischaemic and haemorrhagic stroke and permit the urgent use of thrombolytic drugs in patients with ischaemic stroke. The purpose of this study was to characterize the UCLH Mk 2 MFEIT system, designed for this purpose, with 32 electrodes and a multiplexed 2 kHz to 1.6 MHz single impedance measuring circuit. Data were collected in seven subjects with brain tumours, arteriovenous malformations or chronic stroke, as these resembled the changes in haemorrhagic or ischaemic stroke. Calibration studies indicated that the reliable bandwidth was only 16-64 kHz because of front-end components placed to permit simultaneous EEG recording. In raw in-phase component data, the SD of 16-64 kHz data for one electrode combination across subjects was 2.45 +/- 0.9%, compared to a largest predicted change of 0.35% estimated using the FEM of the head. Using newly developed methods of examining the most sensitive channels from the FEM, and nonlinear imaging constrained to the known site of the lesion, no reproducible changes between pathologies were observed. This study has identified a specification for accuracy in EITS in acute stroke, identified the size of variability in relation to this in human recordings, and presents new methods for analysis of data. Although no reproducible changes were identified, we hope this will provide a foundation for future studies in this demanding but potentially powerful novel application.