Bbg 250

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.

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 dye osmolarities were ca. 298 mOsm, and the pH values were 7.4. We used donor tissue from SightLife (Seattle, WA, USA) for DMEK. All grafts were peeled as described previously. BBG (0.1%, w/v) was used to stain the graft edges during peeling (Figure 1). A punch was gently placed on the endothelial surface to indent a circle 7.75 or 8.0 mm in diameter. Next, 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. 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.