This review summarizes the existing literature on the use of in vitro lung slices to study pulmonary physiology, pharmacology, pathogenesis and toxicity. Since in vitro lung slices maintain cell-cell and cell-matrix relationships in a highly controllable and accessible setting, they offer many advantages over both in vivo and single-cell culture systems. With the advent of high-production slicers, lung slices can be rapidly and reproducibly generated, including from animals treated in vivo. Slices can then be treated in vitro and analyzed using high-throughput technology. Therefore, the lung-slice system offers broad, current and unrealized potential for the detection of toxicity and the delineation of pathophysiologic and therapeutic mechanisms.