颗粒状角膜营养不良活体共焦显微镜形态学研究

目的研究颗粒状角膜营养不良角膜各层组织的共焦显微镜形态改变。方法应用Confoscan2.0共焦显微镜对13例(26眼)颗粒状角膜营养不良患者的角膜进行扫描检查,记录与分析各层角膜图像。结果所有患眼前基质细胞及16/26眼后基质细胞结构不清,排列紊乱,并可见短棒状多形性强反光;6/26眼前弹力层不规则并增厚,神经纤维密度明显下降;6/26眼角膜上皮基底细胞层可见不定型的强反光;2/26眼角膜上皮细胞边界不清,排列呈疏松的蜂窝状,并出现不透明的强反光;所有患者角膜内皮细胞形态基本正常。视力0.3以下的患眼角膜上皮细胞层、上皮基底细胞层、前弹力层、后基质层发生形态异常的比例高于0.3以上的患眼(P<0.05)。结论1.共焦显微镜可活体检查颗粒状角膜营养不良角膜组织各层结构,起到类似病理组织切片的作用。2.前基质层形态异常可能是颗粒状角膜营养不良最基本的共焦显微镜形态特征,病情越重,前基质层以外的其它层次发生形态异常的可能性越大,但内皮细胞层一般不受累。3.共焦显微镜检查对颗粒状角膜营养不良手术方式的选择具有一定的参考价值。     

激光共焦显微镜观察兔眼角膜缘干细胞缺乏羊膜移植后的形态

目的:评价激光共焦显微镜在观察兔眼角膜缘干细胞缺乏后创面修复过程中的作用.方法:角膜缘干细胞缺乏家兔18只(雌雄不限)分A组为正常眼组,B组为角膜缘干细胞缺陷羊膜移植组.两组的家兔于术后第1,3,5,7,10,15,30d进行中央角膜各层组织激光共焦显微镜检查.结果:通过连续共聚焦扫描,获取手术前后角膜各层平面图像(x,y轴)及纵向断层图像(z轴),图像均非常清晰.角膜表皮细胞排列疏松,呈多角型,胞体发暗,边界清晰发亮,明亮的细胞核可见或不可见.术后30d后角膜上皮细胞排列整齐,光反射正常;前基质细胞边缘欠清,胞核明亮呈纺锤形.后基质角膜细胞核较长,相对前基质密度较低.术后30d基质细胞排列紊乱,散在的、反光较强的圆形物质,并产生反射较高的细胞外基质.一些细胞呈梭形,基质产生点状与线形交织的瘢痕组织;基质层内有少量的新生血管,见红细胞在血管内流动;内皮细胞呈规则的六边形,细胞边界呈黑色胞体发亮,看不到细胞核,边缘发暗.术后角膜内皮细胞形态及数量无明显变化.结论:激光共焦显微镜可对角膜进行无创伤、实时活体检查,具有高清晰度、确切的深度定位、时间动态观察、纵向断层扫描等优势,更可提供理想的角膜精确、可重复、深径度图像.     

Chandler综合征的共焦显微镜研究

目的 应用角膜激光共焦显微镜观察Chandler综合征患者活体角膜各层组织的形态特点.方法 对21例(21只眼)的Chandler综合征患者,采用角膜激光共焦显微镜观察其活体角膜的各层结构,总结其图像特点.结果 共焦显微镜下Chandler综合征患者角膜有如下特点:(1)角膜上皮细胞及角膜基质细胞在共焦显微镜下可有不同程度的水肿.(2)角膜营养不良是Chandler综合征患者的特征性改变之一,在共焦显微镜下角膜内皮细胞出现不同程度的疣锥状突起,病变的内皮细胞彻底丧失六边形结构,细胞变圆、变大,在中高反光的细胞体外有一圈由于细胞突起引起的暗反光圈结构,严重的患者可以形成山脊状隆起.(3)部分患者的虹膜在共焦显微镜下可出现粗细不同的支架状结构,这些虹膜支架结构在共焦显微镜下呈中-高度反光,部分呈树枝状排列,部分呈树干状排列,其周围可散在大小不同的色素颗粒附着.结论 共焦显微镜在临床诊断Chandler综合征中有重要作用,角膜营养不良是Chandler综合征患者的特征性改变之一,在共焦显微镜下角膜内皮细胞出现不同程度的疣锥状突起,这种内皮细胞是Chandler综合征与其他类型ICE综合征鉴别的特点之一.     

板层角膜移植术后全层角膜组织改变的共焦显微镜观察

目的研究活体扳层角膜移植术后角膜各层组织的共焦显微镜(confocal microscopy)形态学改变，方法应用Confoscan 2.0共焦显微镜对板层角膜移植术后3月～3年，角膜植片稳定的患者共17例17眼，进行扫描检查，记录与分析各层角膜图像。结果共焦显微镜上能够清楚地观察到稳定角膜的上皮细胞、基底层细胞、前弹力层，它们同正常角膜的相应细胞在图像上没有显著差别。但植片与植床的基质细胞在共焦显微镜图像上有着明显的差异。板层角膜移植术后稳定角膜植片的基质细胞，细胞核较小呈中等反光，排列较植床的基质细胞紊乱。而角膜植床的基质细胞，细胞核较大也呈中等反光，排列较植片的基质细胞稍整齐，结论1、Confoscan 2.0共焦显微镜可活体检查，板层角膜移植术后角膜组织结构和细胞的病理改变。2、板层角膜移植术后可成功的重建眼表。3、活体板层角膜移植术后稳定角膜上有自体角膜基质细胞与异体的角膜基质细胞同时共存，角膜内发细胞的形态结构正常。4、角膜板层间的不平滑与植片基质细胞结构紊乱可能是导致板层角膜移植术后视力不佳的重要原因。     

共焦显微镜观察虹膜角膜内皮综合征及其继发性青光眼的手术治疗

目的应用共焦显微镜检查探讨虹膜角膜内皮综合征的临床特征及其继发性青光眼的手术治疗。方法 8例（8眼）虹膜角膜内皮综合征行共焦显微镜下双眼检查;描述角膜内皮的病变结构特点,分析双眼角膜内皮细胞密度、内皮细胞平均面积、六角形细胞比例及中央角膜厚度,并对6例继发性青光眼中的5例施行手术治疗。结果患眼角膜内皮细胞呈风筝样或上皮细胞样改变;细胞排列紊乱,大小不均;细胞内可见高反光的细胞核,部分可见双核。患眼和对侧眼的角膜内皮细胞密度、内皮细胞平均面积、六角形细胞比例分别为：789.7±75.8个/mm^2、2223.7±80.6个/mm^2;1106.9±89.4μm^2、379.8±20.6μm^2;17.2±1.2%、56.3±1.7%,下降明显。二者的中央角膜厚度相当。4例继发性青光眼手术后眼压控制正常,另1例手术无效。结论共焦显微镜能够发现ICE患者的角膜内皮细胞的特征性改变;对该病的诊断具有很高的临床意义。并发继发性青光眼要及时手术治疗控制眼压。     

三种实验动物角膜活体共焦显微镜检测的比较解剖学研究

背景 海德堡视网膜厚度分析仪和角膜模块的结合实现了对眼表活体组织结构的非侵入性检查,利用共焦显微镜对常用实验动物角膜结构进行比较研究可为相关研究提供依据.目的 利用活体共焦显微镜比较新西兰大白兔、Lewis大鼠、Swiss小鼠的角膜结构,建立实验动物角膜的活体组织图像资料,为共焦显微镜的实验研究提供依据.方法 利用海德堡视网膜厚度分析仪(HRT-Ⅱ)的Rostock角膜模块对新西兰大白兔、Lewis大鼠、Swiss小鼠的角膜进行活体分析,角膜的每一层各采集20张共焦显微镜图片,比较分析实验动物角膜各层的形态学特点及角膜内皮细胞密度.结果 共焦显微镜下3种实验动物角膜表层上皮细胞表现为高反光或低反光的多形细胞,基底上皮细胞表现为暗的细胞质,细胞核不可见,细胞间排列紧密、规则;前弹力层均表现为含有丰富上皮下神经丛的无定形物质.兔角膜基质层在黑色背景中散布着高反光物质,即为角膜基质细胞核,后基质层细胞密度高于前基质层;大鼠和小鼠的角膜基质层仅观察到大量反光的星形结构,无明显的细胞核.3种实验动物的角膜内皮细胞形态相似,均表现为高反光的胞体,边界较暗且细胞排列成蜂窝状.新西兰兔前基质角膜细胞密度中位数为387.5个/mm2,后基质角膜细胞密度中位数为223.5个/mm2,明显少于前基质的细胞密度(U=0.000,P=0.000);新西兰兔、Lewis大鼠、Swiss小鼠角膜内皮细胞密度中位数分别为2192.5、1936.0、1565.0个/mm2,总体差异有统计学意义(H=49.940,P=0.000),兔角膜内皮细胞密度明显高于大鼠和小鼠,差异均有统计学意义(x2=0.000,P=0.000;x2=0.000,P=0.000),大鼠和小鼠的角膜内皮细胞密度差异亦有统计学意义(x2=0.000,P=0.000).结论 共焦显微镜下新西兰大白兔、Lewis大鼠、Swiss小鼠角膜各层的细胞形态相似,但内皮细胞密度和基质细胞形态之间存在明显差异.HRT-Ⅱ的Rostock角膜模块可为动物实验提供角膜各层次的高分辨率图像.     

共焦显微镜在不典型虹膜角膜内皮综合征诊断中应用

目的 探讨共焦显微镜检查在虹膜角膜内皮综合征临床诊断中的应用价值.方法 对8例(其中6例单眼患病,2例双眼患病)常规检查无法确诊的疑似虹膜角膜内皮综合征病例进行共焦显微镜检查.观察角膜内皮层病变结构.结果 活体共焦显微镜检查发现患眼角膜内皮细胞均偏大而且形状不规则,细胞核反光明显增高,可见散在双核、多核、偏位核及分叶核等改变,7例细胞边界模糊,部分病例有局灶内皮细胞缺失、胞内环形暗区、分裂状细胞等改变.此外,3例单眼患病病例的对侧眼在共焦显微镜下发现内皮细胞密度下降、大小不均等改变.最后确诊为原发性进行性虹膜萎缩型2例、Chandler综合征4例、Cogan-Reese综合征2例.结论 共焦显微镜可以在细胞水平上无创、高分辨率地观察到虹膜角膜内皮综合征患眼角膜内皮层特征性的显微结构改变,且不受角膜水肿的影响,大大提高了虹膜角膜内皮综合征诊断的准确性,并有助于早期诊断,具有很高的应用价值.     

共焦显微镜下虹膜角膜内皮综合征的角膜形态学观察

目的 观察共焦显微镜下虹膜角膜内皮综合征(ICE)的角膜各层组织的形态学特点.方法 观察型系列病例研究.对2000年1月至2007年6月在复旦大学附属眼耳鼻喉科医院眼科就诊的23例(23只眼)ICE患者,行共焦显微镜(NIDEK,Confoscan 3.0)下双眼检查.使用NAVIS软件记录和分析角膜各层组织图像,并使用SPSS 13.0软件对患眼的角膜内皮细胞密度、内皮细胞平均面积、六角形细胞比例及有核细胞比例进行单因素方差分析.结果 ICE患眼的角膜内皮细胞呈"风筝样"或"上皮细胞样",细胞形态失去正常的六角形外观,细胞排列紊乱,胞内可见高反光的细胞核,部分细胞内可见双核或细胞分裂象.随病程时间延长,患眼角膜内皮细胞密度和六角形细胞比例逐渐下降.病程短于1年、1～3年、3～5年及超过5年患者的角膜内皮细胞密度分别为(1687.1±122.6)、(1210.6±168.7)、(947.3±145.2)及(856.8±73.4)个/mm2;六角形细胞比例分别为(51.5±6.3)%、(39.8±9.2)%、(32.7±8.1)%及(24.1±5.6)%.随病程时间延长,患眼角膜内皮细胞平均面积和有核内皮细胞比例逐渐升高.病程短于1年、1～3年、3～5年及超过5年患者的内皮细胞平均面积分别为(678.3±56.3)、(928.7±96.2)、(1188.5±72.6)及(1337.5±60.8)μm2;有核细胞比例分别为(12.6±1.4)%、(56.8±3.7)%、(78.7±5.6)%及(84.3±2.8)%(F=7.158,7.736,6.876,14.452;均P=0.000).患眼基质细胞形态无异常,但是随病程延长,基质神经纤维逐渐增粗并明显扭曲.患眼上皮细胞的形态无异常.对侧眼角膜的各层形态结构无明显异常.结论 共焦显微镜下ICE患眼的角膜内皮细胞表现有一定特点,共焦显微镜对于判断ICE的病程进展有诊断价值.     

应用共焦显微镜观察Fuch角膜内皮营养不良患者的病变形态学特征

目的探讨应用共焦显微镜观察我国Fuch角膜内皮营养不良患者角膜各层的活体形态学特征。方法对19例(38只眼)Fuch角膜内皮营养不良患者的中央部角膜进行活体共焦显微镜检查,分为有症状组(19只眼)和无症状组(19只眼),并选取30只眼作为正常对照组,应用NAVIS软件测量、分析角膜各层组织细胞形态和密度,以及滴状赘疣和角膜神经的直径。结果 (1)有症状组:19只眼的角膜内皮层均见到滴状赘疣,直径20-60 μm,内皮细胞密度与正常对照组比较差异有显著意义(t=18.74,P<0.01);9只眼后弹力膜增厚;14只眼角膜后基质层有长条形暗区结构;19只眼角膜基质反光普遍增强;17只眼Bowman膜有局灶性高反光区域;19只眼基底上皮细胞形态大致正常;10只眼显示正常的角膜神经结构;后、前基质细胞密度,与正常对照组比较差异无显著意义(t=0.854、1.173,P=0.38、0.24)。(2)无症状组:19只眼的角膜内皮层均见到滴状赘疣,数目较有症状组者少,直径15-40μm;内皮细胞密度,与正常对照组比较,差异无显著意义(t=1.998,P=0.053);角膜其余各层未见异常。有症状组与无症状组的内皮细胞密度计数比较,差异有非常显著意义(t=8.352,P<0.01)。结论活体共焦显微镜检查有助于Fuch角膜内皮营养不良患者的诊断,特别适用于角膜水肿、角膜内皮镜无法成像的患者。(     

人体角膜移植内皮排斥反应的共焦显微镜研究

目的：应用共焦显微镜探讨活体角膜内皮排斥反应的早期形态学特征和变化。方法：19例(19只眼)角膜移植术后发生内皮排斥反应的人群根据角膜内皮排斥反应轻重的不同分为轻、中、重三组。应用共焦显微镜(Confoscan2．0)对角膜植片进行检查，并分析其相关因素。结果：活体角膜内皮排斥反应特有的共焦显微镜改变主要包括：①早期发现在KP及其附近的内皮细胞面有少量的免疫细胞附着，紧靠KP处的角膜基质细胞出现轻度水肿，细胞排列较混乱，胞浆中出现少量空泡状暗区。②中期发现在内皮细胞表面可见较多的免疫细胞聚集并被完全激活，基质细胞明显水肿，细胞核不清楚，基质中有免疫细胞进入；可见由高反光的免疫细胞和不明成份的组织碎片混合组成内皮排斥线。③在晚期排斥的患者中可见大量“激活”的免疫细胞在角膜内皮细胞面聚集，内皮细胞几乎完全破坏；基质细胞明显水肿，部分被“激活”且伴有大量免疫细胞聚集。结论：①内皮排斥反应的早期能够发现角膜内皮面有免疫细胞沉积；②角膜内皮面的免疫细胞沉积与角膜内皮排斥反应的轻重成正相关；③角膜内皮排斥反应实际上是一个混合排斥反应的过程；④共焦显微镜对角膜内皮排斥反应具有实时、无创、敏感、准确以及可莺蒽、可比较、可活体观察等临床价值。     

分泌减少性干眼症角膜共焦显微镜下的形态学改变

目的应用角膜共焦显微镜活体、无创观察分泌减少性干眼症患者的角膜上皮及上皮下神经纤维的形态学改变。方法选择临床确诊为分泌减少性干眼症的患者37例（73眼）。印迹细胞学检查对患眼进行干燥程度分级,应用角膜共焦显微镜观察角膜上皮下神经纤维的形态学改变,及角膜上皮细胞密度定量测定。结果干眼症患者干燥斑处有明显的角膜上皮细胞坏死脱落区,反光极强;细胞问隙明显增宽,局限上皮细胞缺损、排列稀疏。部分患者呈扁平形浅表层角膜上皮细胞缺失。角膜上皮下神经纤维出现分支紊乱、异常走行弯曲,神经纤维不均匀增粗、增生,7眼未见上皮下神经纤维。干燥程度分级≤Ⅱ级者6眼,角膜上皮细胞密度为（629±168）个／mm^2;Ⅲ级者51眼,细胞密度为（615±181）个／mm^2;≥Ⅳ级者16眼,细胞密度（605±108）个／mm^2,分别与正常眼比较,差异均有统计学意义（P〈0．05）。结论分泌减少性干眼症角膜共焦显微镜下呈现角膜上皮细胞密度降低、上皮下神经纤维的特征性异常改变,与干眼症干燥程度密切相关。     

In vivo corneal confocal microscopy in marfan syndrome

PURPOSE: To examine the cornea of patients with Marfan syndrome in comparison with a control group by using the in vivo confocal microscope. METHODS: Twenty-four eyes of 12 patients with Marfan syndrome had their corneas examined using the in vivo confocal microscope Heidelberg Retina Tomograph (HRT) II/Rostock Cornea Module. The control group included 24 eyes of 12 subjects who had their corneas examined by the same in vivo confocal microscope. RESULTS: Epithelium and neural plexus examination did not show any difference between the 2 groups. Examination of the stroma showed no significant differences concerning the morphology and density of keratocytes. The extracellular matrix of 16 of the 24 eyes of the Marfan group was clearly visible and showed thin highly reflective interconnected lines between keratocytes. In the healthy eye group, reflective lines were observed in only 5 of the 24 eyes. The endothelium of 14 corneas of the Marfan group showed brightly reflective particles. In no cornea of the control group were such particles observed. CONCLUSIONS: Highly reflective extracellular matrix of the stroma and brightly reflective particles among the endothelial cells were the 2 main corneal findings observed by using in vivo corneal confocal microscopy in patients with Marfan syndrome compared with a control group. Further studies need to be made to confirm these findings and eventually find new criteria for Marfan syndrome from the examination of in vivo corneal confocal microscopy.     

激光共焦显微镜对正常人中央角膜的观察

目的:利用激光共焦显微镜观察正常人的中央角膜组织结构和细胞形态。方法:对54例(68眼)患者进行激光共焦显微镜连续扫描,使用激光共焦显微镜对其角膜中央区进行检查,拍摄各层角膜图像,观察组织结构和细胞形态,对细胞密度进行计数并分析。结果:角膜组织学结构:上皮细胞较其它细胞的体积小,数量多,随深度增加细胞逐渐变小。Bowman膜较薄,无明显细胞结构,层间有神经走形。基质层的细胞核围成"网眼"状结构,随深度加深,"网眼"变大,排列变松。Descemet膜表现为一个与后基质层和内皮层交织的移行带。内皮细胞为六边形结构,排列规则。角膜细胞密度:在各年龄组间,上皮细胞表层与基底层之间的差异均有统计学意义(P<0.01),前基质层细胞密度与中、后基质层密度之间有统计学差异(P<0.05)。A组和D组的内皮细胞密度有统计学差异(P<0.05)。其余各年龄段的角膜各层细胞密度,随年龄的增加,数差异均无统计学意义。结论:激光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无创的定性定量分析,较传统光学共焦显微镜能获得更清晰、更密集的角膜图像,对角膜疾病的基础研究与临床诊断具有很高的价值。     

圆锥角膜各期的共焦显微镜表现

目的评价共焦显微镜在圆锥角膜临床研究中的应用价值。方法应用共焦显微镜观察圆锥角膜患者32例（48眼）及正常对照组17例（28眼），分别比较早、中、晚期圆锥角膜与正常对照组的图像特点。结果早期圆锥角膜出现激活状态的角膜细胞、浅基质层的细小皱褶、深基质层的暗纹、部分内皮细胞异形性明显，中、晚期圆锥角膜出现角膜上皮细胞拉伸、细胞核皱缩；基质细胞排列紊乱、基质层暗纹；而对照组未发现上述表现。各期圆锥角膜的角膜基质层厚度、不同深度角膜基质细胞密度、内皮细胞密度与对照组之间差异有统计学意义（P〈0．05）。结论共焦显微镜对早期圆锥角膜的发现以及圆锥角膜病理发展的研究具有重要的临床价值。     

乳化硅油滴在活体角膜内皮的共焦显微镜表现

目的 采用角膜激光共焦显微镜观察乳化硅油滴在活体角膜内皮面的形态,比较并总结其特点.方法 应用自身对照研究.对近年来在深圳市眼科医院就诊的20例(20只眼)临床硅油乳化的患者采用激光角膜共焦显微镜检查其角膜,分析研究其图像特点,总结乳化硅油滴在活体角膜内皮的共焦显微镜表现.结果 乳化硅油滴在角膜内皮面有着不同的大小和排列方式,镶嵌于角膜内皮面,并伴有角膜内皮细胞的明显水肿增大,内皮细胞的结构不清,仅有模糊轮廓.结论 采用新型的激光角膜共焦显微镜可以观察到乳化硅油滴对角膜内皮细胞的直接损害.该检查是一种对人体无害、可以反复进行,并且连续观察角膜内皮变化的工具,对评估乳化硅油滴对角膜的损害具有重要作用.

Abstract：

Objective To study the emulsified silicone oil droplets in the corneal endothelium by in vivo laser confocal microscopy, compare and summarize its characteristics. Methods The corneas of 20 patients (20 eyes) with silicone oil emulsion were examined by in vivo laser confocal microscopy, analyzed the characteristics of the images, and summarized the performance of emulsified silicone oil droplets in corneal endothelium with confocal microscopy. Results Emulsified silicone oil droplets in the corneal endothelium had a different size and arrangement, embedded in the corneal endothelium and come al endothelial cells associated with increased edema; endothelial cell structure was unclear, with only vague outlines. Conlulusions New type of laser corneal confocal microscope can observe the corneal endothelium damage by emulsified silicone oil drops directly. The inspection is a harmless, can be repeated and continuous observation of corneal endothelial changes in the tools. The laser corneal confocal microscope plays an important role on the assessment of silicon emulsion oil droplets on the cornea in the damage.     

A population study of the normal cornea using an in vivo, slit-scanning confocal microscope.

PURPOSE: To document qualitative and quantitative changes in the normal, healthy human cornea with age using the confocal microscope. METHODS: The central corneas of 120 subjects (mean age, 41 years; range, 11 to 80 years) were examined using an in vivo slit-scanning real-time confocal microscope. Images of the corneal stroma and endothelium from both eyes of each subject were semiautomatically analyzed in an observer-masked, randomized manner. RESULTS: Anterior keratocyte density, posterior keratocyte density, and endothelial cell density were shown to be unaffected by the sex of the subject with p values of 0.46, 0.55, 0.50, respectively (multivariate analysis of variance). No statistically significant difference was detected between right and left eyes for all corneal layers examined. The anterior keratocyte density, posterior keratocyte density, and endothelial cell density decreased at a rate of 0.48, 0.22, and 0.33% per year, respectively. A positive correlation was found between the coefficient of cell variation and age. CONCLUSIONS: This data constitutes essential normative data that can be used as a control in further research into abnormal corneal conditions.     

Demonstration of "owl's eye" morphology by confocal microscopy in a patient with presumed cytomegalovirus corneal endotheliitis

PURPOSE: To report confocal microscopic observations of characteristic corneal endothelial lesions in a patient with presumed cytomegalovirus (CMV) corneal endotheliitis. DESIGN: Case report. METHODS: A 77-year-old, immunocompetent man was admitted with corneal edema, keratic precipitates, and coin-shaped lesions in the right eye. Confocal microscopy was performed to examine the corneal endothelium. Polymerase chain reaction (PCR) was used to identify viral DNA in an aqueous humor sample. RESULTS: CMV DNA was detected by PCR. Confocal microscopy showed large corneal endothelial cells with an area of high reflection in the nucleus surrounded by a halo of low reflection. This "owl's eye" morphology is characteristic of CMV infection. Topical and intravenous ganciclovir treatment resulted in rapid resolution of the corneal precipitates and edema, followed by disappearance of the owl's eye morphology. CONCLUSIONS: Confocal microscopy can detect the owl's eye morphology in the corneal endothelium of patients with presumed CMV corneal endotheliitis.     

Comparative anatomy of laboratory animal corneas with a new-generation high-resolution in vivo confocal microscope

PURPOSE: The aim of the current study was to compare the corneas of three commonly used laboratory animals with a new in vivo confocal microscope. METHODS: Six eyes of three adult male New Zealand albino rabbits, six eyes of three adult male Lewis rats, and six eyes of three adult male Swiss mice were used in this study. Corneas were analyzed in vivo using the Rostock Cornea Module of the Heidelberg Retina Tomograph (HRT)-II. For all eyes, 20 confocal microscopic images of each layer, that is, the superficial and basal corneal epithelia, the Bowman layer, the anterior and posterior stroma, and the endothelium, were recorded. The images were then analyzed qualitatively and compared among animals. Cellular densities of anterior and posterior stroma keratocytes of rabbits and endothelium density of the three different animals were also measured and compared. RESULTS: The Rostock Cornea Module of the HRT II was successfully used to analyze all corneal layers of these three commonly used laboratory animals. Although the cellular patterns of the corneal layers of these three animals, as observed with in vivo confocal microscopy, were quite similar, some differences were seen in terms of endothelial cell density and stroma appearance. Superficial cells were seen as hyper- and hyporeflective polygonal cells. Basal cells had dark cytoplasm without visible nuclei and were closely organized. A Bowman layer was observed in all three animals as an amorphous tissue containing fine subepithelial nerve plexus. In rabbits, the stroma consisted of an amorphous ground substance with hyper-reflective structures corresponding with keratocyte nuclei. In rats and mice, numerous reflective stellate structures with no clearly visible nuclei were observed within the stroma. Besides endothelial cell density, the endothelium was similar among the three animals and was seen as hyper-reflective cells with dark limits organized in a honeycomb pattern. CONCLUSIONS: The Rostock Cornea Module of the HRT II can provide high-resolution images of all corneal layers of rabbits, rats, and mice without sacrificing animals or preparing tissue. This new device may be useful for evaluating the cornea during experimental animal studies.     

In vivo laser confocal microscopic analysis of human corneal epithelial sheet cultured on amniotic membrane.

BACKGROUND AND OBJECTIVE: Corneal epithelial sheet cultured on amniotic membrane ex vivo has been developed for the treatment of severe ocular surface disorders. In this study, human corneal epithelial sheets cultured on amniotic membranes were analyzed using a laser scanning confocal microscope. MATERIALS AND METHODS: Human corneal epithelial cells were cultured on four sheets of amniotic membrane, and they were examined layer by layer using the laser scanning confocal microscope. RESULTS: The superficial layer cells were mosaic in appearance. In the superficial layer, focal areas of slender and elongated cells were noted. In addition, highly reflective materials were noted. Basal epithelial cells of the corneal epithelium sheet at a slightly deeper plane displayed mosaic pattern with smaller cells. At a slightly deeper plane, the basement membrane of amnion showed relatively bright homogeneity. CONCLUSION: The laser scanning confocal microscope can examine the morphology of human corneal epithelial cells cultured on amniotic membranes.