Electrical impedance tomography (EIT) visualises alveolar overdistension (OD) and collapse (CL) and enables optimisation of ventilator settings by using the best balance between OD and Cl (ODCL). Besides, the global inhomogeneity index (GI), measured by EIT, may also be of added value in determining PEEP. PEEP is determined based on the best dynamic compliance (Cdyn) without EIT. This study aimed to assess the effect of a PEEP trial on ODCL, GI and Cdyn in patients with and without ARDS. Secondly, PEEP levels from “optimal PEEP” approaches by ODCL, GI and Cdyn are compared. In 2015–2016, we included patients with ARDS using postoperative cardiothoracic surgery (CTS) patients as a reference group. A PEEP trial was performed with four consecutive incremental followed by four decremental PEEP steps of 2 cmH2O. Primary outcomes at each step were GI, ODCL and Cdyn. In addition, the agreement between ODCL, GI, and Cdyn was determined for the individual patient. 28 ARDS and 17 CTS patients were included. The mean optimal PEEP, according to Cdyn, was 10.3 (± 2.9) cmH2O in ARDS compared to 9.8 (± 2.5) cmH2O in CTS patients. Optimal PEEP according to ODCL was 10.9 (± 2.5) in ARDS and 9.6 (± 1.6) in CTS patients. Optimal PEEP according to GI was 17.1 (± 3.9) in ARDS compared to 14.2 (± 3.4) in CTS. Currently, no golden standard to titrate PEEP is available. We showed that when using the GI, PEEP requirements are higher compared to ODCL and Cdyn during a PEEP trial in patients with and without ARDS.