Objective To evaluate the safety and the clinical effects of a single corneal tissue donor specimen split by microkeratome and used for deep anterior lamellar keratoplasty and endothelial keratoplasty.Methods It was a retrospective case series study.Fresh donor corneas were split by microkeratome (Amadeus Ⅱ,Switzerland).The endothelial graft with a small section of the stroma was used for endothelial keratoplasty (EK), and the anterior lamellar button was used for deep anterior lamellar keratoplasty (DALK).Clinical data of 17 consecutive patients who underwent EK from March 2010 to August 2010 and 17 patients who underwent DALK whose donor grafts were matches for the endothelial grafts were reviewed.The attachment of the endothelial grafts and postoperative restitution of the anterior lamellar buttons were evaluated during the 19-24 month follow-up period.Optometric examinations,optical coherence tomography for all patients,and an endothelial cell count for patients who underwent EK were performed.Pre- and postoperative visual acuity analyzed using paired t test.Results The endothelial grafts of the 17 patients who underwent EK had attached well and postoperative visual acuity was improved.The mean astigmatism of the patients was 1.69±0.60 D with the exception of one alkali-burn patient whose optometric data was not available because of corneal opacity.The mean density of endothelial cells for the 17 patients who underwent EK was 2128±244 cells/mm2 and the mean thickness of the endothelial grafts was 152±46 μm.Anterior lamellar buttons of the 17 patients Who underwent DALK were transparent and postoperative best corrected visual acuity improved as well.The mean astigmatism was 2.50±0.90 D and the mean thickness of the anterior lamellar buttons was 343±39 μm.Conclusion The method of splitting a single donor cornea by microkeratome for two recipients is feasible in clinical applications.There were no serious complications caused by donor grafts during the long-term follow-up period. This method,which could make full use of donor corneas and alleviate the current shortage of donor corneas is of great significance.