The invention discloses a square-combination multiple-layered type experimental device for zebrafish capable of simulating crowd flow. The square-combination multiple-layered type experimental devicecomprises a partitioning plate, a hedging cabin, a light supplementing device, a water drainage pipe, a support frame, a carbon-fiber filter, a water purifying box, a base, a turbine water pump, a square fish tank, an oscillation generator, a water inlet pipe, a hemispheric bell and a spring fixer. The square-combination multiple-layered type experimental device disclosed by the invention has thebeneficial effects that a ratio of the length-width size of the square fish tank to the length-width size of a practical crowded building site is 0.08, so that the design of a model is more reasonableand is more comparative; and the zebrafish for simulating the crowd flow, raised in the square fish tank, can realize that group behavior rules of humankind can be reflected more favorably in simulation of emergency evacuation, and a water level can be controlled to about 1cm when the zebrafish is raised in the square fish tank, and action of fish school is controlled to realize strict dimensional action, superposition is avoided, and meanwhile, an influence of a reflection effect caused by depth of the water level on observation also can be reduced.