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To report the study in rabbits of a new method of harvesting Descemet membrane-endothelium complex approaching from the suprachoroidal space for Descemet membrane endothelial keratoplasty.Twenty-four eyes of 12 rabbits were used for microscopic analysis, whereas 24 eyes of 12 cats were used for gross photographic analysis. The enucleated eyeballs of the rabbits and cats were incised 360 degrees circumferentially along the equator. Manual separation was performed between the sclera and the underlying connective tissue approaching from the suprachoroidal space in the direction of the central cornea step by step in spiral pattern. After complete separation of the stroma from the Descemet membrane-endothelium-iris complex, it was excised from the globe at the outside of the iris root. The synthetic carrier (soft contact lens) was laid down on the punch block, and then the excised complex was brought over the carrier with the endothelium-iris side upward. Two steps of down punch for the designed graft size were performed. The harvested complexes were processed for vital stains and were evaluated through photography taken under a light microscope and gross close-up. The endothelial cell loss rates were calculated by a computer software.The overall harvest success rate was 89.6% (43 of 48). The overall structural cell loss rate was 1.83 ± 2.34 by microscopic analysis and 4.68% ± 1.68% by gross photographic analysis.The new method of en bloc harvesting Descemet membrane-endothelium complex approaching from the suprachoroidal space could provide minimal damage to the endothelium during harvesting, and it can be carried and passed through an injector.