The vertical distribution of zooplankton can have substantial influence on trophic structure in freshwater systems, particularly by determining spatial overlap for predator/prey dynamics and influencing energy transfer. The zooplankton community in some of the Laurentian Great Lakes has undergone changes in composition and declines in total biomass, especially after 2003. Mechanisms underlying these zooplankton changes remain poorly understood, in part, because few studies have described their vertical distributions during daytime and nighttime conditions or evaluated the extent to which predation, resources, or environmental conditions could explain their distribution patterns. Within multiple 24-h periods during July through October 2012 in Lake Huron, we conducted daytime and nighttime sampling of zooplankton, and measured food (chlorophyll-a), temperature, light (Secchi disk depth), and planktivory (biomass of Bythotrephes longimanus and Mysis diluviana). We used linear mixed models to determine whether the densities for 22 zooplankton taxa varied between day and night in the epi-, meta-, and hypolimnion. For eight taxa, higher epilimnetic densities were observed at night than during the day; general linear models revealed these patterns were best explained by Mysis diluviana (four taxa), Secchi disk depth (three taxa), epilimnetic water temperature (three taxa), chlorophyll (one taxon), and biomass of Bythotrephes longimanus (one taxon). By investigating the potential effects of both biotic and abiotic variables on the vertical distribution of crustacean zooplankton and rotifers, we provide descriptions of the Lake Huron zooplankton community and discuss how future changes in food web dynamics or climate change may alter zooplankton distribution in freshwater environments.