Recently, it has been discovered that gaseous nickel and iron is present in most comets. To evaluate the state of the laboratory data in support of these identifications, we re-analyzed archived spectra of comet C/1996 B2 (Hyakutake), one of the nearest and brightest comets of the last century, using a combined experimental and computational approach. We developed a new, many-level fluorescence model that indicates that the fluorescence emission of Fe I and Ni I vary greatly with heliocentric velocity. Combining this model with laboratory spectra of an Fe-Ni plasma, we identified 23 lines of Fe I and 14 lines of Ni I in the spectrum of Hyakutake. Using Haser models, we estimate the nickel and iron production rates as Q Ni = 2.6−4.1×10 22 s −1 and Q Fe = 0.4−2.6×10 23 s −1 . From derived column densities the Ni/Fe abundance ratio log 10 [Ni/Fe] = −0.38±0.06 deviates significantly from solar, and it is consistent with the ratios observed in solar system comets. Spectra from several offset distances show profiles consistent with a short-lived parent and/or emissive photodissociation of an unknown parent species. Possible production and emission mechanisms are analyzed in context of existing laboratory measurements. Based on the observed spatial distributions, excellent fluorescence model agreement, and Ni/Fe ratio, our findings support an origin consisting of dissociation of an unknown parent followed by fluorescence emission. Our findings suggest that the strong heliocentric velocity dependence of the fluorescence efficiencies can provide a meaningful test of the physical process responsible for the Fe I and Ni I emission.