A novel technique to covalently immobilize indicator dyes with terminal amino groups for preparing optical sensors is investigated. Au nanoparticles are used as bridges and carriers for anchoring indicator dyes on the surface of a quartz glass slide. 1-Aminopyrene (AP) was employed as an example of indicator dyes and covalently immobilized onto the outmost surface of the glass slide. First, the glass slide was functionalized by (3-mercaptopropyl) trimethoxysilane (MPS) to form a thiol-terminated self-assembled monolayer, where Au nanoparticles were strongly anchored via covalent link. Then, 16-mercaptohexadecanoic acid (MHDA) was self-assembled to bring carboxylic groups onto the surfaces of Au nanoparticles. A further activation by using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) converted the carboxylic group into succinimide ester. Finally, the active succinimide ester was reacted with 1-aminopyrene (AP). Thus, AP was covalently immobilized to the glass slide and an AP-immobilized sensor was obtained. The resulting sensor was used to determine rutin based on fluorescence quenching. It showed a linear response toward rutin (R) from 5.0 × 10-7 to 6.0 × 10-4 mol L-1 with a detection limit of 2.0× 10-7 mol L-1. This AP-immobilized sensor has very satisfactory reproducibility, reversibility, rapid response and no dye-leaching.