Fast 3D live imaging with high resolution has become an indispensable technique in the field of cell and developmental biology to characterize the biological dynamics from the molecular scale up to the entire organism. Nevertheless, live imaging is often not sufficient: precise spatio-temporal manipulation of biological components is required to directly test the underlying mechanisms. Here we present an integrated framework that combines three-dimensional light-sheet imaging in time with precise spatio-temporal photo-manipulations induced by short infrared laser pulses. As a proof of concept, we will use the living Drosophila embryo as a model system and perform in toto imaging in tandem with two-photon optogenetics and multiphoton plasma ablation of the actomyoin network with subcellular precision to test cell and tissue mechanics. Finally, with this hands-on workshop the participants will have the opportunity to learn more about and test on living samples multi-view light sheet microscopy coupled to multiphoton laser manipulation: a powerful tool in modern biology.