Micromixers have been commonly utilized in the areas of biotechnical engineering, analytical chemistry, medical industry, and high-throughput synthesis because biological processes generally involve complicated chemical reactions that require useful reactant mixing for initiation. This paper reports the design, simulation, fabrication and test of a new high-performance micromixer via three-dimensional (3D) Tesla structures to realize the effective mixing process in the microfluidic devices for bio-applications. Both computational fluid dynamics (CFD) calculations and experiments were conducted to study the flow and mixing characteristics in the development of 3D Tesla micromixers. The predicted and measured results of the proposed micromixer have illustrated excellent mixing performance for Reynolds numbers (Re) ranging from 0.1 to 100 (0.015-15 mu L/s) with its pressure drop less than 1054 Pa at Re = 100. We also showed the successful application of the proposed mixer to the immunofluorescence analysis of the EGF receptor L858R mutant specific rabbit mAb (as a cancer biomarker) recognized by anti-rabbit IgG-CFL555 and H1975 cells, revealing the mixing effectiveness of this microfluidic device in the binding reaction of antibodies for detecting antigens of lung cancer cellular surface. (C) 2014 Elsevier B.V. All rights reserved.