Resolution of the nitrogen (N) cycle in the marine environment requires an accurate assessment of dinitrogen (N2) fixation. We present here an update on progress in conducting field measurements of acetylene reduction (AR) and 15N2 tracer assimilation in the oligotrophic North Pacific Subtropical Gyre (NPSG). The AR assay was conducted on discrete seawater samples using a headspace analysis system, followed by quantification of ethylene (C2H4) with a reducing compound photodetector. The rates of C2H4 production were measurable for nonconcentrated seawater samples after an incubation period of 3 to 4 h. The 15N2 tracer measurements compared the addition of 15N2 as a gas bubble and dissolved as 15N2 enriched seawater. On all sampling occasions and at all depths, a 2- to 6-fold increase in the rate of 15N2 assimilation was measured when 15N2-enriched seawater was added to the seawater sample compared to the addition of 15N2 as a gas bubble. In addition, we show that the 15N2-enriched seawater can be prepared prior to its use with no detectable loss (< 1.7%) of dissolved 15N2 during 4 weeks of storage, facilitating its use in the field. The ratio of C2H4 production to 15N2 assimilation varied from 7 to 27 when measured simultaneously in surface seawater samples. Collectively, the modifications to the AR assay and the 15N2 assimilation technique present opportunities for more accurate and high frequency measurements (e.g., diel scale) of N2 fixation, providing further insight into the contribution of different groups of diazotrophs to the input of N in the global oceans.