K L Kratz-Owens1, G S Hageman, D J Schanzlin. 1. Bethesda Eye Institute, Department of Ophthalmology, St Louis University School of Medicine, St Louis, MO 63110.
Abstract
BACKGROUND: We have recently developed a method for labeling and following the migration of keratocytes using the fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI), which intercalates specifically into the adenine-thymine base pairs of deoxyribonucleic acid (DNA). METHODS: DAPI was applied to a 2-millimeter area of the corneal stroma, at the edge of the microkeratome wound, prior to placement of the autograft. RESULTS: DAPI-labeled keratocytes were identified by epifluorescence microscopy along the microkeratome wound and in the mid-periphery of the lamellar graft by day 7. In contrast, no DAPI-labeled keratocytes were observed in the graft immediately following surgery, indicating that the DAPI-labeled keratocytes observed in the graft at day 7 resulted from keratocyte migration, and not from diffusion of DAPI from the host into the graft. CONCLUSIONS: These investigations suggest that DAPI could be a useful probe for monitoring the migration of keratocytes into corneal grafts following lamellar keratoplasty and other corneal transplant surgical techniques.
BACKGROUND: We have recently developed a method for labeling and following the migration of keratocytes using the fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI), which intercalates specifically into the adenine-thymine base pairs of deoxyribonucleic acid (DNA). METHODS:DAPI was applied to a 2-millimeter area of the corneal stroma, at the edge of the microkeratome wound, prior to placement of the autograft. RESULTS:DAPI-labeled keratocytes were identified by epifluorescence microscopy along the microkeratome wound and in the mid-periphery of the lamellar graft by day 7. In contrast, no DAPI-labeled keratocytes were observed in the graft immediately following surgery, indicating that the DAPI-labeled keratocytes observed in the graft at day 7 resulted from keratocyte migration, and not from diffusion of DAPI from the host into the graft. CONCLUSIONS: These investigations suggest that DAPI could be a useful probe for monitoring the migration of keratocytes into corneal grafts following lamellar keratoplasty and other corneal transplant surgical techniques.