Inertial microfluidics offers high throughput, label-free, easy to design, and cost-effective solutions and is a promising technique based on hydrodynamic forces (passive techniques) instead of external ones, which can be employed in lab-on-a-chip and micro-total-analysis-systems for focusing, manipulation, and separation of microparticles in chemical and biomedical applications. The current work, studies the focusing behavior of microparticles in an asymmetric curvilinear microchannel. For this purpose, focusing behavior, including position and band width, of microparticles of diameters of 10, 15 and 20 µm, which served as representatives of different cells, in an asymmetric curvilinear microchannel with curvature angle of 280° was experimentally studied at flow rates from 400 to 2700 µL/min (corresponding to Reynolds numbers between 30 and 205). The results revealed that the largest distance between focusing bands of 20 µm and 10 µm microparticles as well as between focusing bands of 15 µm and 10 µm was obtained at Reynolds number of 121. For the case of microparticles of diameters 20 µm and 15 µm, the largest distance was seen at Reynolds number of 144. The focusing band width became smaller in the asymmetric microchannel so that focusing could be more clearly observed in this configuration.