Comparing the Evaluating Ability of Limbal Insufficiency Using Laser Scanning Confocal Microscopy, Corneal Impression Cytology and Cytospin Assay

1. Wei-Li Chen^1,2
2. Fung-Rong Hu^1,2

¹Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
²Center of Corneal Tissue Engineering and Stem Cell Biology, National Taiwan University Hospital, Taipei, Taiwan

Purpose: To compare the evaluating ability of limbal insufficiency by using laser scanning in vivo confocal microscopy, conjunctival impression cytology, cytospin assay on corneal surface cells and immunohistochemical study.

Methods: Sixteen rabbits corneas of mechanical induced total limbal insufficiency were examined by slit lamp biomicroscopy, laser scanning confocal microscopy, corneal impression cytology with immunohistostaining (K12/K13 double staining staining) on corneal surface cells, cytospin assay of the corneal surface cells cells and immunohistochemcial study.

Results: Although total limbal injury were purposely created and slit lamp biomicroscopic findings implied total limbal insufficiency, the corneal impression cytology and cytospin assay proved that mixed corneal/conjunctival epithelial cells was found in some areas of corneas. In vivo confocal microscopy can provide useful informations in detecting limbal insufficiency (10/16) especially when superficial epithelial layer showed both squamous epithelium and cuboidal small cells. Cytospin assay of corneal surface cells (12/16) is more sensitive than corneal impression cytology (10/16) in detecting limbal insufficiency.

Conclusions:. In vivo confocal microscopy and corneal impression cytology are both non-invasive and can be useful for clinical diagonosis and staging of limbal insufficiency.