Purpose Preloaded, tri-folded grafts in Descemet Membrane Endothelial Keratoplasty require transfer of the tri-folding process from the cornea transplant surgeon to the eye bank technician. We sought to assess if tri-folding may be safely conducted by an eye bank technician with cell loss comparable to standard peeling and lifting. Methods A total of ten grafts were stained, peeled and transferred directly onto a bed of Calcein-AM and Amvisc Plus by an eye bank technician. Five grafts were removed and stained as a scroll and five grafts tri-folded with the endothelium in before transfer. Photographs were acquired with an inverted fluorescence microscope and image segmentation was performed. A t-test was conducted to compare for difference in endothelial cell loss across groups. Results Mean cell loss in the scroll group was 18.5% [95% confidence interval (CI): 15.2-21.9%] compared to 7.6% the tri-folded group (95% CI: 1.7-13.5%). A two-tailed t-test indicated decreased cell loss in the tri-folded group (p=0.013). Conclusion Despite additional manipulation of the graft, tri-folding of Descemet membrane and endothelium may be performed by an eye bank technician without significantly increased cell loss relative to graft preparation as a scroll.