I compared fish assemblages captured in three different microhabitats (shoreline, pelagic near-shore, and benthic near-shore) during day and night fishing in different protection (inside bay or tributary vs. outside in Lake Ontario proper) and turbidity regimes of four near-shore areas of Lake Ontario. The effects of diel movement and availability to gear were clearly evident. Fish assemblages were consistently and significantly more diverse at night than during the day, with nighttime assemblages often being supersets of daytime assemblages. Evidence for a turbidity effect was much weaker than the effects of nocturnal movements and changes in availability to the gear associated with darkness. Nighttime sampling is more likely to capture the full array of species in near-shore areas of the Great Lakes than daytime sampling.