Organotypic brain slice cultures (OSCs) provide a physiologically relevant model for recapitulating complex tissues in vitro. However, the longitudinal monitoring of dynamic processes in OSCs through high-resolution imaging is limited in standard culturing methods, as they rely on slice placement on porous membranes at the air-liquid interface to preserve tissue viability. Here, a new experimental paradigm based on a modular slice culture insert is introduced, which is compatible with static and