The malaria parasite Plasmodium falciparum (Pf) changes the structure and mechanical properties of red blood cells (RBCs). These changes decrease deformability and increase cytoadherence of Pf-infected RBCs to the vascular endothelium, eventually leading to flow occlusions in capillary vessels. In this study, to detect Pf-infected RBCs effectively, deformability and viscosity of blood sample are measured simultaneously and indirectly by quantifying blood flow in a microfluidic device. The microfluidic device is designed by mimicking a Wheatstone-bridge electric circuit. To measure RBC deformability, a deformability assessment chamber (DAC) at the left lower side channel has parallel microfluidic filters. After delivering blood sample and 1× PBS solution at the same flow rate, hemodynamic properties are measured using a time-resolved microparticle image velocimetry technique. Blood volume delivered into the DAC for 200 s is evaluated as a deformability index. Subsequently, blood viscosity is quantified by monitoring blood-filled width of parallel flows in the microfluidic device. The proposed method is applied to evaluate variations in biophysical properties of blood samples partially mixed with normal RBCs and hardened RBCs. As a result, RBC deformability is more effective than blood viscosity in the detection of blood samples with hardened RBC volume fraction of 5%. The microfluidic device is also applied to detect Pf-infected RBCs. When parasitemia is greater than 0.515% for ring stage, 0.0544% for trophozoite stage, and 0.0054% for schizont stage, the measured velocity fields show unstable behavior because of cytoadherence of Pf-infected RBCs. Blood volume delivered into the DAC significantly decreases with increasing parasitemia. The experimental method proposed in this study can detect Pf-infected RBCs with good accuracy.