title: Rapid identification and detection of parasitized human red cells by automated flow cytometry.
abstract: Rapid and reliable identification of various human red cells parasites is important in many chemotherapeutic and immunologic studies. Because manual microscopic counting is tedious and imprecise, we have developed a simple diagnostic procedure for the automated flow cytometric detection of in vitro infected red cells, using a nucleic acid-binding fluorescent dye, acridine orange. Human malaria (Plasmodium falciparum)-infected red cells from continuous human erythrocyte culture were incubated at room temperature in acridine orange stain for 5 min after which the samples were analyzed by flow cytometry. Since mature red cells contain no DNA, infected red cells were identified with a distinct fluorescent signal. A total of 200,000 cells per sample were counted and analyzed in less than 2 min. Rings, trophozoites, and schizonts were assessed and identified in synchronized infected red cell cultures by flow cytometry. In addition, various stages of infected red cells were isolated with a cell sorter. This rapid method permits accurate and reliable assessment of data with the exclusion of anomalous data such as damaged cells, extraneous material, and contaminating particles.
mesh_terms: D000165:Acridine Orange; D002469:Cell Separation; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D004912:Erythrocytes / Q000469:parasitology*; D005434:Flow Cytometry; D006801:Humans; D010274:Parasitology / Q000379:methods*; D010963:Plasmodium falciparum / Q000032:analysis / Q000502:physiology*
chemical_list: D004247:DNA; D000165:Acridine Orange