Four thiophene derivatives were prepared by replacing the 3,4 positions of thiophene by―OCH3, ―CH3―COOCH3 and ―CN, respectively. The polycondensations via direct arylation took place between the four thiophene derivatives and 2,7-dibromo-9,9-dioctylfluorene under six various catalytic conditions to investigate the substituent effects. The substituent can affect the electron cloud density of the active C―H bond, which can be monitored by the NMR chemical shift. The experimental results show that the reactivity decreases with increasing the chemical shift of active C―H bonds in the four thiophene derivatives, and thus can promote the direct arylation polycondensation. This phenomenon is explained by the electrophilic aromatic substitution(SEAr) mechanism. UV-Visible absorption and photoluminescence were studied to investigate the substituent effect on optical properties of the four copolymers. The results show that these alternating fluorene-thiophene copolymers with different substituents are good fluorescent materials and promising in PLED applications. Four thiophene derivatives were prepared by replacing the 3,4 positions of thiophene by-OCH3, -CH3-COOCH3 and -CN, respectively. The polycondensations via direct arylation took place between the fourthiophene derivatives and 2,7-dibromo-9,9 The compound can affect the electron cloud density of the active C-H bond, which can be monitored by the NMR chemical shift. The experimental results show that the reactivity decreases with increasing the chemical shift of active C-H bonds in the four thiophene derivatives, and thus can promote the direct arylation polycondensation. This phenomenon is explained by the electrophilic aromatic substitution (SEAr) mechanism. UV-Visible absorption and photoluminescence were studied to investigate the substituent effect on optical properties of the four copolymers. The results show that these alternating fluorene-thiophene copolymers with differen t substituents are good fluorescent materials and promising in PLED applications.