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Previous animal experiments have suggested that electrical impedance tomography (EIT) has the ability to noninvasively track changes in cardiac stroke volume (SV). The present study intended to reproduce these findings in patients during a fluid challenge. In a prospective observational study including critically ill patients on mechanical ventilation, SV was estimated via ECG-gated EIT before and after a fluid challenge and compared to transpulmonary thermodilution reference measurements. Relative changes in EIT-derived cardiosynchronous impedance changes in the heart (Delta Z(H)) and lung region (Delta Z(L)) were compared to changes in reference SV by assessing the concordance rate (CR) and Pearson's correlation coefficient (R). We compared 39 measurements of 20 patients. Delta Z(H) did not show to be a reliable estimate for tracking changes of SV (CR = 52.6% and R = 0.13 with P = 0.44). In contrast, Delta Z(L) showed an acceptable trending performance (CR = 94.4% and R = 0.72 with P < 0.0001). Our results indicate that ECG-gated EIT measurements of Delta Z(L) are able to noninvasively monitor changes in SV during a fluid challenge in critically ill patients. However, this was not possible using Delta Z(H). The present approach is limited by the influences induced by ventilation, posture or changes in electrode-skin contact and requires further validation.