Fluorescent porous silicon was prepared as a stable biosensor chip substrate. The aminopropyltriethoxysilane (APTES) molecules are attached in the pores of the porous silicon with a crosslink method, and when the molecules are added into the chip, the fluorescence intensity is reduced according to the concentration of the APTES. Controlled experiments are also presented with the small molecule that cannot be covalently coupled, and the results show that this kind of sensor chip has better specificity. Compared with other conventional methods, this method is simple, quick and label- free.