Corneal Graft Marking Technique

BBG 250® (BBG; Sigma-Aldrich, St. Louis, MO, USA) was dissolved in balanced saline solution (BSS® or BSS-plus®; Alcon, Osaka, Japan) to 0.1% (w/v). All grafts were peeled as described previously. BBG (0.1%, w/v) was used to stain the graft edges during peeling. A punch was gently placed on the endothelial surface to indent a circle 7.75, 8.0, or 8.25 mm in diameter. Subsequently, 1.0 and 1.5 mm- diameter dermatological biopsy punches (Kai Industries, Seki, Japan) were used to place asymmetric marks on the edges of the identified circles [19 (link)]. Donor grafts thus marked were stained with 0.1% (w/v) BBG (1.0 mg/mL) for 1 min and stored in BSS prior to insertion, 30 min later [20 (link)]. Because unfolding time depends on donor age, we usually selected donors over 60 years old.