Large river systems create challenges when sampling zooplankton. The dynamics of large rivers, spatial heterogeneity of biota, and behaviours of zooplankton can all contribute to large variation in estimation. The gear utilized for zooplankton collections may also be a factor, and there is not a well‐studied gear, nor industry standard, for collection in riverine systems. The lack of consistent sampling methods makes it difficult to compare zooplankton within and across systems. A standard method for zooplankton collection would be valuable for comparison among studies. To optimize zooplankton assessment in large river habitats, we tested how community estimations varied between four common zooplankton sampling devices across three river habitat types. We tested four gears: the Schindler–Patalas trap, integrated tube sampler, powered water pump, and horizontal tow net. Each device was used to collect samples in thalweg, channel border, and backwater river habitats within Pool 8 of the Upper Mississippi River in June, July, and August of 2017. Our results support that there are qualitative and quantitative differences in zooplankton estimates among gear in different habitats. The powered pump most often yielded highest abundances of total and individual zooplankton taxa allowing for more reliable community comparisons. With some modification, the pump is recommended as the most appropriate sampling gear when performing quantitative studies of zooplankton composition and abundance in large river habitats.