Purpose: To establish an in vitro model of laser in situ keratomileusis (LASIK) in human donor eyes and to test its validity in comparison with animal models. Setting: Department of Anatomy, Friedrich-Alexander Unviersity, Erlangen, Germany. Methods: Laser in situ keratomileusis was performed on 20 organ-cultured human corneal buttons. The excimer laser ablations ranged from 0 to 12.0 diopters. The corneas were maintained in culture for up to 6 months and then evaluated with light microscopy and transmission electron microscopy. In addition, corneal sections were immunohistochemically stained for collagen type III, laminin, and fibronectin. The main outcome measures were the ultrastructural and immunohistochemical features of the stromal incision interface. Results: Ultrastructural investigations in the peripheral cornea revealed a disarrangement of collagen fibers, indicating scar formation. These findings were not observed in the central area. Immunohistochemical staining for fibronectin and collagen type III was detected over the entire stromal incision interface, whereas laminin staining was related to the ingrowth of epithelial cells. Conclusions: The morphological changes after LASIK in an organ culture model can simulate the in vivo situation. Therefore, this model appears appropriate to use in further study of corneal wound-healing changes after LASIK.