A method for separation and staining of flat mounts of human corneal endothelium

A technique is described in which sheets of corneal endothelium are removed from human donor corneo-scleral discs. Celloidin solution was applied to the endothelial surface, allowed to dry, peeled off with the attached endothelial cell layer and mounted on a glass slide. Following removal of the celloidin with acetone, this endothelial cell flat mount was then stained with H&E and monoclonal antibodies to cell adhesion molecules. A pilot study of endothelial cell adhesion molecule expression in flat mount preparations of 14 corneas showed constitutive neural cell adhesion molecule (NCAM) expression, but a lower degree of focal expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VACM)-1, P/E-selectin and HLA-DR.