Isolation of microparticles and biological cells on microfluidic chips has received considerable attention due to their applications in numerous areas such as medical and engineering fields. Microparticles separation are of great importance in bioassays owing to the need for a smaller sample and device size, and lower manufacturing costs. In this study, we first explain the concepts of separation and microfluidic science along with their applications in the medical sciences, and then, a conceptual design of a novel inertial microfluidic system is proposed and analyzed. The PDMS spiral microfluidic device was fabricated, and its effects on the separation of particles with sizes similar to biological particles were experimentally analyzed. This separation technique can be used in the process of separating cancer cells from the normal ones in the blood samples. These components required for testing were selected, assembled, and finally, a very affordable microfluidic kit was provided. Different experiments were designed, and the results were analyzed using appropriate software and methods. Separator system tests with polydisperse hollow glass particles (diameter 2–20 µm), and monodisperse Polystyrene particles (diameter 5,15 µm), and the results exhibit an acceptable chip performance with 86 percent of efficiency for both monodisperse particles and polydisperse particles. The microchannel collects particles with an average diameter of 15.8 µm, 9.4 µm, and 5.9 µm at the Proposed reservoirs. This chip can be integrated into a more extensive point-of-care diagnostic system to test blood samples.