Suspended particles are an essential component of large rivers influencing channel geomorphology, biogeochemical cycling of nutrients, and food web resources. The Upper Mississippi River is a large floodplain river that exhibits pronounced spatiotemporal variation in environmental conditions and biota, providing an ideal environment for investigating dynamics of suspended particles in large river ecosystems. Here we investigated two questions: (i) How do suspended particle characteristics (e.g. size and morphology) vary temporally and spatially? and (ii) What environmental variables have the strongest association with particle characteristics? Water sampling was conducted in June, August, and September of 2013 and 2014 in Navigation Pool 19 of the Upper Mississippi River. A FlowCAM® (Flow Cytometer and Microscope) particle imaging system was used to enumerate and measure particles 53–300 μm in diameter for size and shape characteristics (e.g. volume, elongation, and symmetry). Suspended particle characteristics varied considerably over space and time and were strongly associated with discharge and concentrations of nitrate + nitrite (NO₃⁻) and soluble reactive phosphorus. Particle characteristics in backwaters were distinct from those in other habitats for most of the study period, likely due to reduced hydrologic connectivity and higher biotic production in backwaters. During low discharge, phytoplankton and zooplankton made up relatively greater proportions of the observed particles. Concurrently during low discharge, concentrations of chlorophyll, volatile suspended solids, and total phosphorus were higher. Our results suggest that there are complex interactions among space, time, discharge, and other environmental variables (e.g. water nutrients), which drive suspended particle dynamics in large rivers.