Prognostication of numerous chronic diseases are in need of identifying circulating tumor cells (CTCs), afterwards, separating and reviving contaminated samples are required. Conventional methods of separating blood cells, namely cytometry or magnetically activated cell sorting, in many cases lose their functionality, or efficiency under different conditions. Hence microfluidic methods of separation have been implemented. Herein, an innovative integrated double stair-shaped microchannel is designed and optimized, capable of ‘separation’, and ‘chemical lysis’ simultaneously in which the lysis reagent concentration can be controlled to tune the lysis intensity. The method of insulator-based dielectrophoresis (iDEP), which is the main physics in this device, is utilized yielding maximum separation. Pivotal features of the applied voltage, the voltage difference, the angles and the number of stairs, and the width of the throat in the microchannel have been numerically explored in order to optimize the channel in terms of separation and the lysis buffer concentration. The overall state of optimum case for the voltage difference (ΔV) of 10 owns the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 μm, and the inlet voltages are 30 V and 40 V. Also, the overall state of optimum cases for delta possess the following features: the number of stairs is 2, the angle of stairs is 110°, the width of throat is 140 μm, and the inlet voltages are 30 V and 35 V.