Corneal histopathology is essential to understanding the mechanisms of
corneal disease and wound healing. We have developed a light microsco
pic trichrome stain for 1-mum, aldehyde-fixed, osmicated, epon/resin-e
mbedded corneal sections. The stain consists of azure II, methylene bl
ue, and basic fuchsin. This combination of stains provides a fast and
effective method to clearly differentiate corneal cells from their sur
rounding extracellular matrix, and chromatically separates various ECM
materials within the cornea. Correlative examination by light and tra
nsmission electron microscopy (TEM) provides multilevel understanding
of corneal morphology and ultrastructure. Tissues examined using this
staining procedure may be subsequently examined by TEM for ultrastruct
ural morphology. This stain has proved beneficial in our research of b
oth cellular and extracellular constituents of the cornea.