For many years, application of RBC indices has been recommended for discriminating between subjects with iron deficiency from those with thalassemia. However, application of the algorithms resulted in only 30% to 40% of subjects being appropriately classified. The aim of the study was to establish the efficacy of algorithms for anemia screening including new hematologic parameters such as percentage of hypochromic and microcytic RBCs and hemoglobin content of reticulocytes. Subjects with iron deficiency anemia (IDA) (n = 142) and subjects with β-thalassemia (n = 34) were enrolled in a European multicenter study. Apparently healthy subjects were used as a reference group (n = 309). Hemocytometric investigations were performed on a Sysmex XE5000 hematology analyzer. The algorithms for IDA discrimination yielded results for area under the curve, sensitivity, specificity, and positive and negative predictive values of 0.88, 79%, 97%, 74%, and 98%, respectively. The algorithms for β-thalassemia discrimination revealed similar results (0.86, 74%, 98%, 75%, and 99%, respectively). We conclude that the advanced algorithms, derived from extended RBC parameters provided by the Sysmex XE5000 analyzer, are useful as laboratory anemia screening devices.