Cardiac output (CO) and stroke volume (SV) are parameters of key clinical interest. Many techniques exist to measure CO and SV, but are either invasive or insufficiently accurate in clinical settings. Electrical impedance tomography (EIT) has been suggested as a noninvasive measure of SV, but inconsistent results have been reported. Our goal is to determine the accuracy and reliability of EIT-based SV measurements, and whether advanced image reconstruction approaches can help to improve the estimates. Data were collected on ten healthy volunteers undergoing postural changes and exercise. To overcome the sensitivity to heart displacement and thorax morphology reported in previous work, we used a 3D EIT configuration with 2 planes of 16 electrodes and subject-specific reconstruction models. Various EIT-derived SV estimates were compared to reference measurements derived from the oxygen uptake. Results revealed a dramatic impact of posture on the EIT images. Therefore, the analysis was restricted to measurements in supine position under controlled conditions (low noise and stable heart and lung regions). In these measurements, amplitudes of impedance changes in the heart and lung regions could successfully be derived from EIT using ECG gating. However, despite a subject-specific calibration the heart-related estimates showed an error of 0.0 ± 15.2 mL for absolute SV estimation. For trending of relative SV changes, a concordance rate of 80.9% and an angular error of -1.0 ± 23.0° were obtained. These performances are insufficient for most clinical uses. Similar conclusions were derived from lung-related estimates. Our findings indicate that the key difficulty in EIT-based SV monitoring is that purely amplitude-based features are strongly influenced by other factors (such as posture, electrode contact impedance and lung or heart conductivity). All the data of the present study are made publicly available for further investigations.