Detection of phosphatidylserine-expressing red blood cells in whole blood

There are various mechanisms of red blood cell (RBC) clearance from the circulation, including eryptosis and hemolysis. The spleen plays a major role in eliminating RBCs from the blood flow thanks to their ability to pass through narrow spaces. Still, some questions such as the role of phosphatidylserine-binding proteins in this process remain unanswered. Thus, RBC elimination mechanisms are still not very well explored. Here, we report on a flow cytometry protocol using phosphatidylserine markers for the detection of whole blood erythrocytes that can be classified as pre-elimination RBCs. We also provide the results of healthy donors' blood testing suggesting that reliable detection of eryptosis in whole blood is indeed possible. The study was approved by the Independent Ethics Committee and the Scientific Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of Russia.