[A study on the viability of corneal endothelium in rabbit by dual staining with trypan blue and alizanin red S].

PURPOSE

We study the viability of preserved rabbit corneal endothelium by dual staining with trypan blue and alizanin red S.

METHOD

We randomly divided New Zealand white rabbit eyes into four groups, each with 5 eyes. The eye balls were preserved in moist chambers with aqueous remorad and C3F8 temponade for 5, 7, 10 and 14 days. Then we got the corneal Bottons with sclera ring. Isolated corneas were placed with the endothelial side up in a corneal cup, and trypan blue (0.25%) was added drop-wise to cover the endothelium. After 1.5 minutes the stain was poured out. The endothelial layer was then covered with aliznin red S (0.2%) for 1.5 minutes and pouring away the staining reagent. After staining, the corneas were immensec in glutaraldehyde fixative solution for 10 minutes. Finally, the endothelium was examinated and taken photoes with light microscope.

RESULTS

Changed cells that have become permeable to trypan blue show deep blue staining of their nuclei in contrast to the unstained condition of normal cells. All the intercellular borders of endothelial cells were purple. The survival rate of the endothelium preserved for 5 and 7 day is more than 95%, for 10 days is 75-80%, and for 14 days about 70%.

CONCLUSION

This technique allows a proportion cout of damaged and undamaged cells in the endothelial monolayer, makes it possible to observe their structures clearly and gives advantage to take pathelage photos.