Davison P F, Galbavy E J
Invest Ophthalmol Vis Sci. 1986 Oct;27(10):1478-84.
The fluorescent dye dichlorotriazinyl aminofluorescein will bind to amino groups of proteins covalently under physiological conditions. It has been used to dye the connective tissue around an ulcer or nonpenetrating, linear incision in the rabbit cornea and sclera, and the healing of the tissue has been examined up to 1 yr later. Sagittal sections were stained for light microscopy, and adjacent unstained sections were examined in the fluorescence microscope. The stained sections showed the reestablishment of the lamellar organization of the stromal collagen across the site of the incision; the fluorescence showed where the connective tissue that was present when the wound was made persisted, and thereby defined the limits of remodeling in the healing process. In Bowman's layer and the adjacent stroma, there was often an abrupt transition from fluorescent to new, undyed connective tissue. Deeper in the scar, and particularly around the ulcer, dark streaks were observed between the fluorescent lamellae, showing apparently new (non-fluorescent) tissue interdigitating with the old. These observations are discussed in relation to the mechanism of healing and the residual mechanical weakness that persists across scars in the cornea and sclera.
荧光染料二氯三嗪基氨基荧光素在生理条件下会与蛋白质的氨基共价结合。它已被用于对兔角膜和巩膜溃疡或非穿透性线性切口周围的结缔组织进行染色,并在长达1年后检查组织的愈合情况。矢状切片进行苏木精-伊红染色用于光学显微镜观察,相邻的未染色切片则在荧光显微镜下检查。染色切片显示切口部位基质胶原的板层结构得以重建;荧光显示伤口形成时存在的结缔组织持续存在的位置,从而确定了愈合过程中重塑的范围。在Bowman层和相邻基质中,荧光结缔组织与新的未染色结缔组织之间常常存在突然的过渡。在瘢痕深处,尤其是溃疡周围,在荧光薄片之间观察到深色条纹,表明明显有新的(非荧光)组织与旧组织相互交错。结合愈合机制以及角膜和巩膜瘢痕处持续存在的残余机械性薄弱情况对这些观察结果进行了讨论。