The increased utility of fluorescence-based methods in recent years has highlighted the need for brighter, more efficient fluorophores. In order to design these fluorophores, an improved fundamental understanding is necessary of the structural components that intrinsically effect fluorescence efficiency. Here, we characterize the intrinsic effects of deuteration on fluorescence from gaseous oxazine dyes, without the influence of dye-solvent interactions, by making use of an ion trap mass spectrometer that has been altered to enable optical measurements. Comparison of emission spectra of four oxazine dyes: cresyl violet, oxazine 4, oxazine 170, and darrow red, show little change in profile upon deuteration of amine groups. However, deuteration significantly increases the efficiency of fluorescence with an increase in fluorescence lifetime and brightness by 10-23% for the gaseous dyes. This increase is less than half that of the quantum yield increase observed in deuterated solution. This indicates the large fluorescence efficiency changes for the oxazine dyes in deuterated solution result from a combination of both intrinsic effects as well as substantial contribution from altered fluorophore-solvent interactions. The intrinsic effects behind increased lifetime upon deuteration are explored using time-dependent density functional theory (TD-DFT) calculations of potential energy surfaces (PESs) for ground and low lying excited electronic states. In accord with experimental observations, calculated S1-S0 emission spectra show only minor differences between deuterated and non-deuterated forms indicating that the deuteration does not affect the radiative channel appreciably. Relaxed PES scans along the torsional motions of the amino groups reveal that the increase in lifetimes upon deuteration is likely due to quenching of different radiationless changes channels in different oxazine dyes. Calculations suggest that tunneling to access twisted intramolecular charge transfer states in S1 is critical in several of the oxazines. However, in at least one of the dyes examined, the large isotope effect is more likely due to differences in intersystem crossing rates. Overall, this combined experimental and computational investigation elucidates the photophysics of a well-known fluorescent scaffold and provides insight into how small differences can dramatically affect fluorescence outcomes.