应用共聚焦显微镜观察正常老年人角膜形态学的特点

In vivo morphologic characteristics of healthy old chinese cornea by confocal microscopy

目的:探讨正常老年人共焦显微镜下角膜各层组织的活体细胞形态学特征。方法:正常老年人19例22眼中央角膜应用共聚焦显微镜(confocal microscope through focus,CMTF)进行观察,记录上皮翼状细胞层、上皮基底细胞层、前基质层、后基质层、内皮层的细胞密度,选取每人上皮基底细胞层下神经丛和中后基质层神经最清晰图像,并记录此幅图像中神经纤维总长度、神经纤维直径、神经纤维数目和每100μm神经纤维包含的念珠状结构(beads)数目,同时比较不同眼别及不同性别之间上述计量和计数资料之间的差别;观察上皮翼状细胞层、上皮基底细胞层、前基质层、后基质层、内皮层及上皮基底细胞层下神经丛(subbasal epithelial nerve plexus)和中后基质层神经(stromal nerves)的组织形态。结果:上皮翼状细胞层、上皮基底细胞层、前基质层、后基质层、内皮层的细胞密度分别为2150±315,5270±539,859±137,627±184,2529±654个/mm2,上皮基底细胞层下神经丛每幅图像中神经纤维总长度、神经纤维直径和数目、每100μm神经纤维包含的念珠状结构分别为944±176μm,2.3±0.5μm,9.0±1.3条,4.9±1.4;中后基质层神经每幅图像中神经纤维总长度、神经纤维直径和数目、每100μm神经纤维包含的念珠状结构分别为306±138μm,5.6±1.7μm,1.8±1.5条,0.0±0.0。上述计量和计数资料左、右眼和男、女之间无统计学差异。同时观察到排列疏松、细胞间夹杂有无结构暗区的表层上皮细胞层,过度形态的翼状上皮细胞层,呈"斑马皮样"外观的基底细胞层,在角膜中后基质层有4眼(18%)可见到微皱褶(microfolds),在排列规则、多呈六边型的角膜内皮层发现有2眼(9%)出现类似滴状赘疣(pseudoguttata-like)结构。结论:共焦显微镜可以在实时、活体和三维空间从细胞水平对角膜各层结构进行定量和定性分析。

AIM. To investigate the cellular morphologic characteristics of healthy old chinese cornea by confocal microscopy in vivo. METHODS： Nineteen patients＇（22 eyes） central cornea were examed by confocal microscopy in vivo. The cell densities of wing epithelial cell layer, basal epithelial cell layer, anterior stromal layer, posterior stromal layer and endothelial cell layer were counted. Nerve fibres＇whole lenth, diametre, number of nerve fibres and beads number per 100μm nerve fibre in single clear picture in sub-basal epithelial nerve plexus and in mid-posterior stromal nerve fiber were counted. The differences of these numerical datas between the left and the right eyes and between man ＂s and women ＂s eye were analyzed. Also, the morphologic characteristics of epithelial cell layer, stromal layer and endothelial cell layer were investiaated. RESULTS. The cell densities of wing epithelial cell layer, basal epithelial cell layer, anterior stromal layer, posterior stromal layer and endothelial cell layer were 2 150 ±315, 5 270±539,859 ±137,627 ±184, and 2 529 ±654 cells/mm^2 respectively. The nerve fibres＇whole lenth, dia- metre, number of nerve fibres and beads number per 100 m nerve fibre in single clear picture in sub-basal epithelial nerve plexus were 944 ± 176μm, 2.3 ± 0.5μm, 9.0 ± 1.3 pieces and 4.9 ± 1.4 respectivley. The nerve fibres＇whole lenth, diametre, number of nerve fibres and beads number per 100μm nerve fibre in single clear picture in mid-posterior stromal nerve fiber were 306 ± 138μ m, 5.6 ± 1.7μm, 1.8 ± 1.5 pieces, and 0.0 ± 0.0 respectively. The differences of these numerical datas between the left and the right eyes and between man＇s and the women＇s eyes had no significance. The superfical epithelial cell layer appeared loosing arrangement with no-structure areas scattered. The wing epithelial cell layer showed cellular transition characteristics. The basal eoithelial cell arrangement looked like ＂ girrafe skin＂. CMTF images displayed microfolds （4 eyes） in the mid-posterior stroreal layers and pseudoguttata-like changes （2 eyes） in the well arranged hexagon endothelial cell layer. CONCLUSION. In vivo CMTF examination can offer qualitative and quantitative analysis in real-time and 3D condition to all layers of cornea in cellular level. It is a useful method to reveal corneal morphologic changes both physiologically and pathologically.