Objective To evaluate the effect of CO 2 -insufflation with 5 and 10 mmHg on cerebral oxygenation and hemodynamics in neonates. Background An increasing percentage of surgical interventions in neonates are performed by minimal invasive techniques. Recently, concerns have been raised regarding a decrease of cerebral oxygenation in neonates during thoracoscopy as a result of CO 2 -insufflation. Methods This was an animal experimental study. Piglets were anesthetized, intubated, ventilated, and surgically prepared for CO 2 -insufflation. Insufflation was done with 5 or 10 mmHg CO 2 during 1 h. Arterial saturation (SaO 2 ), heart rate (HR), mean arterial blood pressure (MABP), and cerebral oxygenation (rScO 2 ) were monitored. CFTOE, an estimator of cerebral oxygen extraction ((SaO 2 -rScO 2 )/ SaO 2 )), was calculated. Arterial blood gases were drawn every 15 0 : pre (T0), during (T1-T4) and after CO 2 -insufflation (T5). Results Ten piglets (4 kg) were randomized for 5 (P5) and 10 (P10) mmHg CO 2 -insufflation. Two P10 piglets needed resuscitation after insufflation, none P5. Linear mixed-effect modeling of paCO 2 , pH, and SaO 2 showed that values were dependent on time and time squared (p \ 0.001) but were not different between the 5 and 10 mmHg groups. Analysis demonstrated significant changes over time in heart rate and MABP between the 5 and 10 mmHg groups, with a significant higher heart rate and lower blood pressure in the 10 mmHg group (p \ 0.001). For rScO 2 and cFTOE, no group differences could be demonstrated, but a significant effect of time was found: rScO 2 increased and cFTOE decreased (p \ 0.001). Conclusions Insufflation of CO 2 during thoracoscopy with 10 mmHg caused more severe hemodynamic instability and seems to be related with a decrease of cerebral perfusion as represented by a higher oxygen extraction. CO 2 -insufflation of 5 mmHg for thoracoscopy seems to have no adverse effects on cerebral oxygenation.