Polyfluorene and its derivatives (PFs) are extremely appealing electrochemiluminescence (ECL) illuminants thanks to their easy modification, high quantum yield, excellent photostability, and nontoxicity, exhibiting great application potential in ECL sensing and imaging. Unfortunately, most reported PFs-based ECL bioanalysis generally exhibited high triggering potential (>1.0 V vs Ag/AgCl), which introduced undesirable electrochemical interference to adversely affect the sensitivity and accuracy of biological analysis. This work innovatively exploited poly(3,4-ethylenedioxythiophene) (PEDOT) as an interfacial conductor to modulate the low ECL triggering potential of poly[(9,9-dioctylfluorenyl-2,7-diyl)-