A two-dimensional micro-fluidic detection technique was studied. Micro-fluidic device designed distinguishingly was fabricated through methods of laser ablation, laser lithography and chemical etching. Continuous laser is used to excite fluorescence from the microprobes passing the detection area and molecules reaction information of suspension array is obtained by using highly sensitive cooling CCD freezing imaging technology. Two-dimensional micro-fluidic detection technique of suspension array, with high sensitivity and speediness and nicety, was validated by experiment.