Incorporating conjugated polymers with functionalities to provide good adhesion to solid substrates is favorable to prepare fluorescent solid materials for various applications. Inspired by the wide range and robust adhesion property of mussel, a series of conjugated polymers with the poly[p-(phenylene ethynylene)-alt-(thienylene ethynylene)] backbone and pendant catechol groups via Sonogashira coupling polymerization were designed and synthesized. The contents of catechol groups were precisely tuned through copolymerization, followed by selective deprotection. For comparison, similar polymers with pendant phenol groups were also synthesized. The conjugated polymers were demonstrated to have expected chemical structures via systematic characterizations. All the polymers had good solubility in common organic solvents and displayed strong fluorescence in solution. To evaluate the adhesion performance of different polymers, fluorescent silica microspheres were selected as the substrate for loading the abovementioned polymers via immersing the silica microspheres into solutions of conjugated polymers. To find out the loading amount, a calibration curve for each polymer was set up based on the linear relationship between the fluorescence intensities of the suspension of microspheres and the polymer concentrations. The loading amounts of catecholfunctionalized polymers were significantly higher than those of phenol-functionalized polymers; the highest loading amount is over 14-fold of the amount of the control polymer without any catechol/phenol functionality. By combining the abovementioned results with the preliminary mechanism investigation based on the solvent washing, it was concluded that catechol functionality and its content in the polymers greatly affected the adhesion performance. The multidentate interaction provided by catechol groups may enable the conjugated polymers to be widely used for fluorescence coating and staining of a variety of substrates.