Underwater strobe lights can influence the behavior and distribution of fishes and are increasingly used as a technique to divert fish away from water intake structures on dams. However, few studies examine how strobe lights may affect organisms other than targeted species. To gain insight on strobe lighting effects on nontarget invertebrates, we investigated whether underwater strobe lights influence zooplankton distributions and abundance in Lake Oahe, South Dakota. Zooplankton were collected using vertical tows at 3 discrete distances from an underwater strobe light to quantify the influence of light intensity on zooplankton density. Samples were collected from 3 different depth ranges (0–10 m, 10–20 m and 20–30 m) at <1 m, 15 m and ⩾100 m distance intervals away from the strobe light. Copepods represented 67.2% and Daphnia spp. represented 23.3% of all zooplankton sampled from 17 August to 15 September 2004. Night time zooplankton densities significantly decreased in surface waters when strobe lights were activated. Copepods exhibited the greatest avoidance patterns, while Daphnia avoidance varied throughout sampling depths. These results indicate that zooplankton display negative phototaxic behavior to strobe lights and that researchers must be cognizant of potential effects to the ecosystem such as altering predator–prey interactions or affecting zooplankton distribution and growth.