Descemet-stripping Automated Endothelial Keratoplasty (DSAEK): Effect of Nontoxic Gentian Violet Marking Pen on DSAEK Donor Tissue Viability by Using Vital Dye Assay

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To qualitatively assess the endothelial damage on Descemet-stripping automated endothelial keratoplasty (DSAEK) donor tissue resulting from a gentian violet marking pen.


An in vitro model was used, by using corneoscleral rims, DSAEK quality corneal donor tissue, and a gentian violet marking pen. After making a mark on the stromal side of a microkeratome-prepared DSAEK corneal button to confirm appropriate orientation of the donor tissue after insertion into the anterior chamber, the corneal tissue was returned to Optisol GS solution for 1 hour. A vital dye assay was used to identify devitalized and necrotic endothelial cells with alizarin red S and trypan blue.


Corneal donor tissue evaluated with the gentian violet marking pen showed positive trypan blue staining, limited to the area marked with the gentian violet ink.


Marking the DSAEK donor stromal surface with a gentian violet marker damages the endothelium. Surgeons should limit the size of the mark or use an insertion technique that avoids confusion about orientation of the donor cornea.

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