活体共焦显微镜下观察正常人眼球结膜组织结构

摘要

目的 探讨应用活体激光共焦显微镜观察正常人球结膜的组织结构.方法 横断而研究.2008年2月至7月选择50名无眼部外伤、感染及配戴接触镜史,且裂隙灯显微镜检查无异常的正常人的50只眼作为研究对象.使用激光共焦显微镜对其上方、下方、鼻侧及颞侧球结膜进行检查,各层图像均被记录,分析球结膜各层形态,并对上皮细胞、杯状细胞及树突状细胞密度进行计数.运用单因素方差分析对各层上皮细胞密度和各方位杯状细胞密度进行统计学分析,运用最小显著性差异分析组间差异.结果 球结膜上皮浅表层细胞体积较大,排列松散,胞核呈低反光,细胞平均密度为(1643±206)个/mm2.中间层细胞呈卵圆形,体积较小,排列紧密,胞核呈点状高反光,细胞平均密度为(4693±228)个/mm2.基底层细胞呈多边形,排列规整,有清晰而高亮的细胞边界,细胞平均密度为(4420±230)个/mm2.经统计学分析3种上皮细胞密度存在显著件差异(F=1160.312,P=0.000).类似杯状细胞的细胞呈圆形,胞体较大,胞内充满透亮颗粒,成团或散在分布,细胞的平均密度为(432±72)个/mm2.树突状细胞呈高反光颗粒,伴树枝状突起,分散于结膜各层,细胞的平均密度为(22±25)个/mm2.在结膜上皮与固有层之间,存在一层致密的高反光的基底膜.球结膜固有层由高度血管化的疏松结缔组织组成,可见大量小规则的条状纤维或大片的网状纤维,弥散分布小圆周高亮的游走细胞,还可清晰观察到血管中血液的流动.结论 活体激光共焦显微镜是研究球结膜组织结构的一种有效上具,为眼表疾病的临床诊断提供了快速而无创的检查手段.%Objective To analyze the morphology of human bulbar conjunetiva by in vivo laser scanning confocal microscopy. Methods This research was a cross-sectional study. From February to July 2008, 50 eyes of 50 healthy subjects were enrolled in this study. They had no history of ocular trauma, infection or contact lens wear and had no found after routine slit-lamp examinations. In vivo laser scanning confocal microscopic examinations were performed on the superior, inferior, nasal and temporal bulbar conjunctiva and the images were recorded. The morphology of bulbar conjunctiva was analyzed and the density of epithelial cells, dendritic cells and goblet cells were calculated. One-way analysis of variance (ANOVA) was used to compare the means of epithelial cell densities in different layers and goblet cell densities in different positions. Subsequently the datum between two groups were analyzed by least significant difference (LSD). Results Superficial epithelial cells of bulbar conjunctiva were characterized as large loose-arranged cells with a hyporeflective nucleus. The mean density is (1643±206) cells /mm2.Intermediate epithelial cells were captured with features of oval small tight-arranged cells with a punctiform hyperrcflective nucleus. The mean density is (4693±228) cells/mm2. Basal epithelial cells appeared to be polygonal and regular-arranged within hyperreflocfive cell borders. The mean density is (4420±230) cells/mm2.There was a significant difference among three kinds of conjunctival epithelium (F = 1160.312,P =0.000). The presumed goblet cell was defined as a large hyperreflective oval-shaped cell with relatively homogeneous brightness, crowded in groups or mainly dispersed. The mean density is (432±72) cells /mm2. The dendritic cell appeared to be hyperreflective corpuscular particles with dendritic processes scattered among conjunctival epithelial cells. The mean density is (22±25) cells /mm2. The basement membrane, a prominent hyperreflective band, separated epithelial cells from subepithelial structure. Bulbar conjunctival substantia propria, beneath the basement membrane, was mainly composed of highly vascularized, loose connective tissues which were irregularly arranged fibers or a network of fibers, punctiform hyperreflective immune cells and sharp flows of blood vessels. Conclusion In vivo laser scanning confocal microscopy is a useful tool in the analysis of the bulbar conjunctival morphology, which provided a fast and noninvasive method for the diagnosis of ocular surface diseases.