Extracorporeal membrane oxygenation (ECMO) can be lifesaving but suffers from high rates of bleeding and repeated transfusions. Current monitoring of blood cell damage during ECMO is limited to platelet counts, hematocrit, and plasma hemoglobin levels. Extracelluar vesicles (EV) are small cell fragments released when cells are activated/injured. The objective was to evaluate flow cytometric measurements of EV during ECMO as an indication of platelet, red cell, and endothelial activation/injury. Samples were collected from 55 patients (1 day to 19 years) during 58 ECMO runs. ECMO activated or injured blood cells, but the extent was highly variable and patient dependent. On average platelet activation was increased sevenfold during ECMO with up to 60-fold increased activation during the first 24 hours in some patients. EV associated with platelet and red-cell injury were increased eightfold on average but up to 200-fold in patients with disseminated intravascular coagulation, severe hemolysis, or massive transfusion. Approximately 9% of ECMO patients showed a red-cell and endothelial activation pattern that was associated with poor prognosis. Extracellular vesicles with autofluorescence similar to bilirubin appeared to come from monocytes processing hemoglobin. ECMO is associated with a highly variable, sustained increase in platelet, red-cell, and endothelial activation and injury that is a combination of circuit and transfusion related events, the patients underlying condition and possibly genetic influences on blood cell activation and injury. Extracellular vesicle measurements may improve our understanding of cellular activation and injury during ECMO as we work to improve the biocompatibility of these systems.