A holographic three-dimensional multi-spot light stimulation device is provided with: a three-dimensional imaging holographic optical system A which employs fluorescent exciting light to acquire three-dimensional fluorescence distribution information resulting from fluorescent signal light from a plurality of stimulation target objects; and a three-dimensional light stimulation holographic optical system B which employs a light stimulation hologram generated on the basis of the acquired three-dimensional fluorescence distribution information to form a plurality of light spots in space, to impart stimulation simultaneously to the plurality of stimulation target objects. Furthermore, the three-dimensional light stimulation holographic optical system B is provided with a spatial light phase modulating element 22 and a control unit 25, wherein the control unit 25 generates the light stimulation hologram by controlling the spatial light phase modulating element 22 on the basis of the three-dimensional fluorescence distribution information.