A series of donor-acceptor (D-A) co-oligomers with oligo(fluorene-alt-bithiophene) and perylene diimide as donor and acceptor segments, respectively, have been designed and synthesized. They can self-assembly into alternating D-A lamellar nanostructured films with the periods depending on the molecular length. These films have been successfully used in fabrication of high-performance single-molecular solar cells with power conversion efficiency up to 1.50%.