Milton Keynes MK7 6AA, United Kingdom ABSRACT 2',3',5'-Tri-O-acetyl-6,8-dithioguanosine (taDTGuo) is an analog of nucleoside and currently under investigation as a potential agent for photodynamic therapy (PDT). Excitation by simultaneous two-photon absorption of visible or near-infrared light would provide an efficient PDT for deep-seated tumors. The two-photon absorption spectrum of taDTGuo was obtained by optical-probing photoacoustic spectroscopy (OPPAS). A two-photon absorption band corresponding to the S 5 ←S 0 transition was observed at 556 nm, and the two-photon absorption cross section (2) was determined to be 26±3 GM, which was much larger than those of other nucleobases and nucleosides. Quantum chemical calculations suggested that the large (2) value of taDTGuo was responsible for large transition dipole moments and small detuning energy resulting from the thiocarbonyl group at 6-and 8-positions. This is the first 1