背景激光共焦显微镜可从细胞水平对正常和病变组织进行活体观察,在眼表系统中的应用越来越广泛,但国内对正常球结膜的活体激光共焦显微镜检测结果报道较少.目的利用活体激光共焦显微镜观察分析正常球结膜的形态.方法应用德国海德堡激光共焦显微镜(HRT3)对健康志愿者15例21眼的上方、鼻下方、鼻侧、颞侧球结膜进行扫描,分辨率为1μm,激光波长为670 nm.观察视野为400μm×400μm.分析球结膜细胞的形态,对球结膜上皮细胞进行计数,并计算杯状细胞和树突状细胞(DC)的密度.结果球结膜表层上皮细胞胞核小,边界模糊.基底上皮细胞边界清晰晰但胞核不可见.类似杯状细胞的细胞呈卵圆形,胞体较大,细胞内充满高反光均一明亮颗粒.成群或散在分布,上皮表面的孔洞结构可能是已经分泌排出内容物的杯状细胞.DC呈高反光颗粒,伴树枝状突起,分布于球结膜上皮各层.球结膜基质由密集的白色网状纤维构成,其间可见含有细胞成分的血管穿过.球结膜表层上皮细胞密度和基底上皮细胞密度分别为(2556±692)mm2和(2985±376)mm2,杯状细胞和DC的密度分别为(77±39)mm2和(26±35)mm2.经统计学分析各方位上皮细胞密度差异无统计学意义(P=0.204,P=0.130),各方位杯状细胞和DC的差异有统计学意义(P=0.001,P=0.000).结论激光共焦显微镜是活体研究人类球结膜形态的有用工具,有助于眼表疾病的诊断和治疗.%Background Application of confocal microscopy in the investigation of ocular surface system in living eye have been greatly extended in two decades.In vivo confocal microscopy allows the evaluation of the normal or pathological tissue at the cellular level.However,seldom study about the morphology of normal in vivo human bulbar conjunetiva under the confocal microscopy was induced. Objective Present study was to analyze the appearance of normal bulbar conjunetiva cells under the in vivo confocal microscopy. Methods Laser scanning confocal microscopy was used to examine the morphological characteristics of the bulbar conjunetiva in 21 eyes of 15 healthy volunteers.The parameters of confocal microscopy were as follows:resolution 1 μm,wave length 670 nm,field range 400 μm×400 μm.The epithelial cell numbers,dendritics cells density and goblet cells density from superior,inferonasal,nasal and temporal bulbar conjunctiva were calculated respectively and compared and imaged by the HRT3 Rostock Cornea Modual. Results The superficial epithelial cells of the bulbar conjunctiva was seen with the small cell nuclei and blur border.The borders of basal epithelial celt were clearly visible without cell nuclei.The presumed goblet cell presented with a large hyperreflective oval-shaped cell with relatively homogeneous brightness,crowed in groups or mainly dispersed.The orfices at the epithelial surface represented the goblet cells,showing some open and expel contents.The dendritic cell appeared to be hyperreflective corpuscular particles with visible processes among conjunetival epithelial cells.A few dense white fiber meshwork was exhibited in conjunctival stroma with the traverse blood vessels containing cellular elements.The superficial and basal epithelial cell densities were 2556±692and 2985±376 cells/mm2 respectively,and overall goblet and mature dendritic cells densities were 77±39 and 26±35 cells/mm2 respectively.Significant differences were proved in globet cell density and dendritic density among different conjunetival zones(P=0.001,P=0. 000),however,the alteration of conjunetival epithelium cells was insignificant in different area(P=0.204,P=0.130).Conclusion Confocal microscopy is a useful tool for the study of morphology of human bulbar conjunctiva cells in vivo.which offer a valuable aid in the diagnosis of ocular surface disease.
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