Near infrared spectroscopy (NIRS) has been used to evaluate regional cerebral tissue oxygen saturation (ScO₂) during the last decades. Perioperative management algorithms advocate to maintain ScO₂, by maintaining or increasing cardiac output (CO), e.g. with fluid infusion. We hypothesized that ScO₂ would increase in responders to a standardized fluid challenge (FC) and that the relative changes in CO and ScO₂ would correlate. This study is a retrospective substudy of the FLuid Responsiveness Prediction Using Extra Systoles (FLEX) trial. In the FLEX trial, patients were administered two standardized FCs (5 mL/kg ideal body weight each) during cardiac surgery. NIRS monitoring was used during the intraoperative period and CO was monitored continuously. Patients were considered responders if stroke volume increased more than 10% following FC. Datasets from 29 non-responders and 27 responders to FC were available for analysis. Relative changes of ScO₂ did not change significantly in non-responders (mean difference − 0.3% ± 2.3%, p = 0.534) or in fluid responders (mean difference 1.6% ± 4.6%, p = 0.088). Relative changes in CO and ScO₂ correlated significantly, p = 0.027. Increasing CO by fluid did not change cerebral oxygenation. Despite this, relative changes in CO correlated to relative changes in ScO₂. However, the clinical impact of the present observations is unclear, and the results must be interpreted with caution. Trial registration:http://ClinicalTrial.gov identifier for main study (FLuid Responsiveness Prediction Using Extra Systoles—FLEX): NCT03002129.