Jumbo multiple plate samplers were suspended in a river at 0.3 and 1 m depth in one of three orientations: interplate spaces closed to downwelling light and open to flow, open to light and flow, or open to light and closed to flow. Mean numbers of colonizing taxa and individuals were not significantly different among orientations after eight weeks of submergence. All variables tested decreased significantly with depth. Mean number of taxa, number of individuals (1 m only), and insect diversity decreased between samplings at four and eight weeks for samplers that were closed to light and open to flow. All orientations, depths, and durations had one‐half to two‐thirds of the total taxa. Colonization was affected by location in the river and position on sampler suspension equipment. The results indicate lack of orientation effects on colonization or high variability that obscured such effects. The sampler suspension equipment possibly increased among‐sampler variability by forming artificial snag habitats, and interplate light and flow conditions at different orientations may not have been sufficiently distinct to elicit different biological responses. Individual samplers provided diverse microhabitats regardless of orientation, but it would be prudent to include orientation among the variables considered in use of multiple plate samplers.