An in-vivo technique for monitoring keratocyte migration following lamellar keratoplasty.
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.
Kratz-Owens
K L
KL
Bethesda Eye Institute, Department of Ophthalmology, St Louis University School of Medicine, St Louis, MO 63110.
Hageman
G S
GS
Schanzlin
D J
DJ
D000818:Animals; D002465:Cell Movement / Q000502:physiology; D003319:Corneal Stroma / Q000473:pathology*; D016039:Corneal Transplantation; D005456:Fluorescent Dyes; D007211:Indoles / Q000008:administration & dosage; D008856:Microscopy, Fluorescence; D011817:Rabbits; D014945:Wound Healing / Q000502:physiology
D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI