8-Vinyladenosine (8VA) is an adenosine analog, like 2-aminopurine (2AP), that has a red-shifted absorption and high fluorescence quantum yield. When introduced into double-stranded DNA (dsDNA), its base-pairing and base-stacking properties are similar to those of adenine. Of particular interest, the fluorescence quantum yield of 8VA is sensitive to base stacking, making it a very useful real-time probe of DNA structure. The fundamental photophysics underlying this fluorescence quenching by base stacking is not well understood, and thus exploring the excited state electronic structure of the analog is warranted. In this study, we report on changes in the electronic structure of 8VA upon optical excitation. Stark spectroscopy was per-formed on 8VA monomer in frozen ethanol glass at 77 K to obtain the direction and degree of charge redistribution in the form of the difference dipole moment, Delta(mu) over bar (01), = 4.7 +/- 0.3 D, and difference static polarizability, tr(Delta(paired right arrows)alpha(01)) = 21 +/- 11 angstrom(3), for the S-0 -> S-1 transition. In addition, solvatochrormsm experiments were performed on 8VA in various solvents and analyzed using Bakhshiev's model. High level ab initio methods were employed to calculate transition energies, oscillator strengths, and dipole moments of the ground and excited states of 8VA. The direction of Delta(mu) over bar (01) was assigned in the molecular frame for the lowest optically accessible state. Our study shows that the angle between ground and excited state dipole moment plays a critical role in understanding the change in electronic structure upon optical excitation. Compared to 2AP, 8VA has a larger difference dipole moment which, with twice the extinction coefficient, suggests that 8VA is superior as a two-photon probe for microscopy studies. To this end, we have measured the ratio of the two-photon fluorescence yields of the two analogs by excitation at the respective monomer absorption maxima. We show that 8VA is indeed a significantly brighter two-photon fluorophore, based on our experimental and computational results.