This technical note describes a facile technique to screen some anticancer drugs and evaluate their effects on nonadhesive leukemic cells in an easily fabricated microfluidic device by utilizing the Annexin V conjugated quantum dots as apoptosis detection probes. The cell immobilizing structures and gradient-generating channels were integrated within the device which was fabricated in one-single step. The nonadhesive leukemic HL-60 cells can be felicitously immobilized and cultured on the dam structures at a proper lateral pressure. We then delivered Annexin V functionalized quantum dots which can readily bind to the outer membrane of apoptotic cells and distinguish the apoptosis from unaffected cells with single cell level resolution. The diffusion time of quantum dots reduced to 5 min before imaging. The capabilities of evaluating drug effect on HL-60 cell line have been shown in both population way and individual cell level. The technique presented herein can bridge the gap between the quantum dots based in vitro cell imaging and the analysis of individual apoptotic cell in a microfluidic system, allows an easy operating protocol to screen some clinically available anticancer drugs.