The validation of UV reactors is currently accomplished by biodosimetry, in which the reactor is treated as a "black-box" and hence cannot account for the dependence of the dose delivery on the complex hydrodynamics and the spatial variation in UV intensity. Alternative methods, such as fluorescent microspheres as non-biological surrogates and computational fluid dynamics (CFD) simulations, have been developed; however, each method has its shortcomings. In this study, a novel technique for the spatial and temporal assessment of the hydrodynamics and the UV dose delivered and the link between these two factors in a lab-scale UV reactor using three dimensional laser induced fluorescence (3DLIF) is developed. This tool can also be utilized for the optimization of UV reactors and to provide data for validation of CFD-based simulation techniques. Regions of optimization include areas around the UV lamp where short-circuiting occurred, a longer inlet approach section that enhances the performance of the reactor by reducing short circuiting paths and a longer outlet region to provide greater mixing. 3DLIF allows real time characterization of mixing and dose delivery in a single lamp UV reactor placed perpendicular to flow by capturing fluorescence images emitted from a laser dye, Rhodamine 6G, using a high speed CCD camera. In addition to three-dimensional mixing, the technique successfully visualized the two-dimensional, transient mixing behaviors such as the recirculation zone and the von Karman vortices and the fluence delivery within the reactor, which has not been possible with traditional tracer test techniques. Finally, a decomposition technique was applied to the flow and fluence delivery based concentration data to reveal similar structures that affect these phenomena. Based on this analysis, changing the flow in the reactor, i.e. the Reynolds number, will directly affect the fluence delivery.