High-throughput microsphiltration to assess red blood cell deformability and screen for malaria transmission–blocking drugs

摘要: The mechanical retention of rigid erythrocytes in the spleen is central major hematological diseases such as hereditary spherocytosis, sickle-cell disease and malaria. Here, we describe use microsphiltration (microsphere filtration) to assess erythrocyte deformability hundreds thousands samples parallel, by filtering them through microsphere layers 384-well plates adapted for discovery compounds that stiffen Plasmodium falciparum gametocytes, with aim interrupting malaria transmission. Compound-exposed gametocytes are loaded into plates, filtered then transferred imaging analysis. High-content detects viable upstream downstream from filters quantifies spleen-like retention. This screening assay takes 3-4 d. Unlike currently available methods used red blood cell (RBC) deformability, enables high-throughput pharmacological (tens tested a matter months) involves challenge induces physiologically relevant dumbbell-shape deformation. It therefore directly assesses ability RBCs cross inter-endothelial splenic slits vivo. protocol has potential applications quality control transfusion determination phenotypic markers diseases.