A convenient method for the synthesis of tris(2,2’-bipyridyl) dichlororuthenium(Ⅱ)hexahydrate-doped amino-modified double-layer silica nanoparticles is presented in this paper.The synthesized nanoparticles are uniform and photostable,and can be well dispersed in a water solution.Proteins could be directly immobilized onto these nanoparticles by a simple coupling process without losing their biological activities.These nanoparticles were further used as fluorescent probes in protein microarray assay for the quantitative detection of protein.The results obtained by these nanoparticles,with the detection limit of as low as 3.5μg/mL,were much better than those involving the use of conventional FITC probe.