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The manual dissection technique for deep lamellar endothelial keratoplasty (DLEK) surgery is technically difficult and may not be smooth enough for consistently optimal postoperative vision. We evaluated the feasibility and efficacy of using a femtosecond laser to perform the dissections in the DLEK procedure.The Intralase femtosecond laser (with standard LASIK surgery spot settings) was used to create a 9.4-mm wide, 400-μm deep lamellar pocket dissection and a 5.0-mm wide side cut near-exit incision in 10 “recipient” whole cadaver eyes and in 10 “donor” cadaver corneal-scleral caps mounted onto an artificial anterior chamber. Recipient and donor disks were resected with special scissors, and the donor tissue was transplanted using the small incision (5.0-mm) DLEK technique. Topography of the recipient eyes was measured pre- and postlaser dissection, and the recipient and donor tissues were sent for scanning electron microscopy (SEM) analysis of the smoothness of the dissections.Successful lamellar dissections were obtained in all tissues. The mean recipient topographic corneal curvature postoperatively was 43.3 ± 1.7 diopters, which was not a significant change from the preoperative curvature of 44.0 ± 0.8 diopters (P = 0.430). The mean recipient topographic astigmatism postoperatively was 1.7 ± 0.8 diopters, which was not a significant change from the preoperative recipient astigmatism of 1.6 ± 0.7 diopters (P = 0.426). Comparison of the histology of the laser-formed stromal dissections by scanning electron microscopy, however, did not appear significantly better than histology after manual DLEK dissections in either the recipient or the donor tissues.A femtosecond laser can create the lamellar dissections for the DLEK procedure, making this procedure easier and faster. As in the manual technique, corneal topography is unchanged by this surgery. More work will need to be done, however, to optimize the laser settings to provide even smoother interface surfaces.