The integration of aggregation-induced emission (AIE) and two-photon absorption (2PA) into one molecule is very important because such a combination would endow the material with a synergetic effect for diverse and broader applications. Herein, two regioisomers based on perylene diimide and anthracene moieties are designed and synthesized. The as-prepared compounds exhibit distinct AIE and 2PA properties, where the bay-substituted one exhibits common AIE and 2PA properties, while the ortho-substituted one exhibits not only enhanced and accelerated AIE performance, but also improved 2PA properties. This research would provide a general approach to explore efficient molecules by the marriage of accelerated AIE and enhanced 2PA that could improve their performance in various areas. We present an isomerism strategy to study the substitution position of perylene diimides on the performances of aggregation-induced emission (AIE) and two-photon absorption (2PA). Interestingly, enhanced AIE and 2PA have been achieved.