The miniaturization of conventional immunoassays using microfluidic techniques has proven promising in bringing accurate and quantitative clinical diagnosis to the point-of-care (POC). However, the microfluidics community has yet to produce many commercial devices for such applications. The main reason for this is the difficulty in producing low-cost, rapid, sensitive, automated, and portable immunosensors. To achieve this goal, our group at the University of Rhode Island has focused on studying the following interrelated scientific and engineering challenges: (1) minimization of the manufacturing cost, (2) analysis time reduction, (3) sensitivity enhancement, (4) automated reagent handling, and (5) miniaturization of the detection system. The potential of using the developed technologies in clinical applications (e.g. immunoglobulin G detection and cardiac risk assessment) is also presented. Ultimately, the developed microfluidic diagnostic device will be extended to perforin rapid, sensitive, and automated diagnosis for a variety of disease biomarkers and virulent pathogens at the POC.