Supravital and Vital Staining of Diseased Corneal Endothelium in Whole-Mount Preparations

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Abstract
• Whereas specular microscopy may provide valuable preoperative information about the status of the endothelial cells in clear corneas, in many diseased corneas that come to penetrating keratoplasty, stromal opacities or epithelial edema prevents determinations about the condition of the endothelium. A method has been developed wherein diseased-host penetrating keratoplasty specimens are maintained postoperatively in McCarey-Kaufman medium and are thereafter stained as whole mounts with alizarin red S and trypan blue to evaluate the status of the endothelial monolayer. Seventy diseased corneas were studied with this method. The alizarin red S stained the intercellular junctions, while the trypan blue stained nuclei and provided information about the viability of the endothelial cells that were studied. Unusual findings were the presence of binucleated endothelial cells in decompensated cases of iris clip pseudophakos and scattered iris stromal melanocytes on bared Descemet's membrane in cases of chemical injury, trauma, and congenital glaucoma. The method was found to be rapid, reliable, and inexpensive, particularly when compared with alternative methods of evaluation, such as scanning electron microscopy.