The 2,7-bis(4-pyridyl)fluorene (BPF) was synthesized via a Suzuki coupling reaction. The optical spectra properties of BPF and BPF-deoxyribonucleic acid (DNA)-cetyltrimethyl ammonium (CTMA) thin films composed of BPF, DNA, and CTMA were characterized by the measurements of UV/Vis absorption spectra and fluorescence spectra. The amplified spontaneous emission (ASE) of the BPF-DNA-CTMA films was researched experimentally by pumping of a pulse laser with a wavelength of 355 nm. The results show that the absorption peak and the fluorescence peak of BPF are located at 327 and 380 nm, respectively. The emission peak of BPF corresponds to the vibronic transitions from an excited state of level to the ground state of level. The ASE peak of the BPF-DNA-CTMA film is located at 384 nm, and the threshold of ASE excited energy density is .