Currently the two most commonly configurations of optical microscopy used in neuroscience laboratories are wide field illumination and laser scanning imaging. In wide field microscopy, the whole field of view is simultaneously illuminated, allowing fast image acquisition or fast repetitive stimulation, but preventing the application of complex spatial light patterns (fig. 1A). Differently, in laser scanning microscopy a diffraction limited laser spot is sequentially deflected in the field of view, allowing the selective illumination of portions of the sample that depend on the scanning trajectory. This configuration leads to an increase of the spatial but to a significant loss in the time resolution of the optical system. Both approaches thus have intrinsic limitations with respect to the degree of complexity with which spatio-temporal patterns of light can be projected onto the biological sample. Illumination with structured light represents an innovative alternative to overcome these limitations. In this experimental approach, the laser wavefront is engineered (or shaped) to simultaneously and selectively illuminate only specific regions of interest in a given field of view. This technique offers flexibility in the pattern of illumination that cannot be achieved with more traditional optical approaches and gives the opportunity of imaging/stimulating simultaneously multiple portions of a given cell or different cells within a neuronal network, or the possibility to manipulate simultaneously different object to controll the micro-enviroment around a cell.