共焦显微镜对近视人群中央角膜组织学改变与年龄相关性的研究

目的探讨共焦显微镜对近视人群中央角膜各层组织的活体观察和分析。方法选择2003年8月-2007年12月天津医科大学眼科中心进行屈光手术术前检查者122例（122眼）,其中男54例（54眼）,女68例（68眼）;年龄18～80岁,平均（28．54±12．4）岁;等效屈光度数为-0．50～-6．0D。检查并记录各层角膜图像,并对各层细胞形态、细胞密度进行分析。结果角膜全层厚度、基质厚度与年龄均呈负相关（r1=-0．552,P=0．014;r2=-0．545,P=0．035）。上皮层基底膜细胞密度与年龄呈负相关（r=-0．355,P=0．017）。前基质、后基质细胞密度与年龄均呈负相关（r1=-0．462;P=0．001;r2=-0．403,P=0．016）。内皮细胞密度与年龄呈负相关（r=-0．603,P=0．006）。随着年龄的增长,角膜内皮细胞发生了形态学改变,内皮细胞的多形性百分比增大,六边形细胞数的百分比下降（r1=0．417,P=0．004;r2=-0．598,P=0．002）。结论近视人群中央角膜组织随着年龄的增长,角膜上皮基底膜细胞、角膜基质细胞、角膜内皮细胞数量下降,角膜全层、角膜基质厚度减少。     

应用共聚焦显微镜检查近视眼中央角膜的临床分析

共聚焦显微镜（confocal microscopy through focusing, CMTF）是近年来发展起来的一种高清晰度、高放大倍率的活体显微镜检查技术,可显示角膜细胞形态和数量变化,直接测得角膜上皮、基质及角膜全层厚度.与传统的角膜组织学标本固定染色方法相比较,在活体状态下,共聚焦显微镜能够无创伤、实时地从细胞学水平观察角膜各层细胞形态结构和动态变化,为研究提供客观、定量的测量数据.     

活体共聚焦显微镜检查在角膜营养不良诊断中的应用

活体共聚焦显微镜检查(in vivo confocal microscopy,IVCM)无创、快速,可活体观察眼表组织及细胞结构的生理、病理变化,在感染性角膜炎诊断及治疗评价、干眼病眼表评估、眼表手术治疗效果评估及糖尿病早期筛查等方面均有广泛应用。利用IVCM观察角膜营养不良有助于诊断、监测疾病进展、评价治疗效果及了解其病理生理变化等。     

Ⅱ型糖尿病患者角膜的共聚焦显微镜观察

目的共聚焦显微镜下观察Ⅱ型糖尿病患者角膜上皮下神经纤维以及角膜各层细胞密度和形态学改变。设计比较性病例系列。研究对象选取确诊为Ⅱ型糖尿病的患者20例(20眼)为病例组和就诊于眼科门诊的无糖尿病的老年性白内障患者20例(20眼)为对照组。方法病例组和对照组均进行角膜共聚焦显微镜检查,对所得的图片和结果进行分析。主要指标角膜上皮下神经纤维密度、角膜上皮基底细胞密度、角膜浅基质细胞密度、角膜中基质细胞密度、角膜深基质细胞密度、角膜内皮细胞密度、内皮细胞的变异系数和六角形百分比。结果糖尿病组与对照组相比,上皮下神经纤维密度明显减少(t=-4.951,P=0.000);角膜浅基质细胞密度、中基质细胞密度和深基质细胞密度均有显著下降(P值均=0.000);角膜内皮细胞变异系数增加(t’=3.652,P=0.001),六角形百分比减少(t=-3.778;P=0.001)。结论共聚焦显微镜下显示Ⅱ型糖尿病患者角膜上皮下神经纤维密度和各层细胞密度下降。     

活体共聚焦显微镜在角膜病临床研究中的新进展

活体共聚焦显微镜能在细胞水平实时、非侵入性、高清晰地检测角膜结构,它已广泛应用于角膜病的研究.本文对活体共聚焦显微镜在感染性角膜炎、圆锥角膜、角膜后沉着物、长期应用抗青光眼药物引起的角膜病变及糖尿病相关的角膜病变的临床研究新进展进行综述.     

《活体角膜激光共聚焦显微镜图谱》一书出版

由北京同仁眼科中心、北京市眼科研究所孙旭光教授主编《活体角膜激光共聚焦显微镜图谱》一书,已由人民军医出版社出版。活体角膜激光扫描共聚焦显微镜分辨率1μm,放大倍率800倍,通过对角结膜组织无创、实时的观察,使眼科医生在活体上,从细胞水平直接观察角结膜及其众多疾病的组织细胞学变化,动态了解疾病的发展和转归,深入探讨疾病的病理机制。本图谱收集整理了作者及其课题组多年来积累的典型角结膜病例,包括感染性角膜病、免疫性角膜病、角膜营养不良与变性、神经营养性角膜病变、角膜手术后、药源性角膜病变、干眼、全身疾病相关性角膜病以及外伤和肿瘤等,通过300余幅图片,展现和总结了共聚焦显微镜在临床指导角结膜病诊断及治疗方面的应用经验。     

共焦显微镜对活体Francois角膜营养不良中央混浊的观察

目的：报告用共焦显微镜观察活体Francois角膜营养不良中央混浊的结果。方法：2位不相关的患者，一位78岁男性和一位75岁女性。均患有Francois角膜营养不良伴中央混浊，用常规裂隙灯活体显微镜和共焦显微镜进行检查。     

共聚焦显微镜下LASIK术后角膜基质的变化

目的研究准分子激光原位角膜磨镶术（LASIK）术后角膜基质的改变。方法选择LASIK患者24例（48眼），于术后第1、10、30和90d进行角膜中央部位共聚焦显微镜检查。结果48眼（100％）角膜瓣基质有微皱褶，在瓣与基质床界面处可见高反光颗粒。界面上下可见激活细胞，激活细胞所在区域深度与角膜瓣厚度呈负相关性（r1=-0．554，P=0．047）；与削切深度呈正相关（r2=0．559，P=0．010），后基质细胞术后有所增加，1个月（P=0．000）时达到最高，3个月时又下降，但仍高于术前（P=0．000）。结论LASIK术后角膜基质可见微皱和高反光颗粒，基质细胞被激活，界面两侧基质中出现无细胞区。后基质细胞密度有所增加。     

共聚焦显微镜在活体角膜上皮肿瘤鉴别诊断中的应用

目的 通过活体共聚焦显微镜在角膜上皮肿瘤诊断中的应用,分析、鉴别角膜上皮肿瘤性质.方法 7例角膜上皮肿瘤患者术前均行常规眼部裂隙灯检查、眼前节OCT和活体共聚焦显微镜检查.所有患者均实施角膜肿瘤切除术,术后行肿瘤组织病理学检查,与术前共聚焦显微镜结果比较.结果 7例角膜上皮肿瘤均侵犯透明角膜、角膜缘和球结膜,突起于眼球表面,形状似桑葚,血管丰富.在眼前节OCT检查中5例角膜肿瘤显示密度均匀,与角膜组织之间界限清晰,浅层基质中未见肿瘤密度影;另外2例角膜肿瘤显示,密度不均,与角膜组织界限不清,而且侵犯前弹力层和浅层角膜基质.在活体共聚焦显微镜中7例均显示角膜上皮细胞非典型增生,其中2例非典型增生明显,侵犯前弹力层,在角膜上皮层和浅基质层内形成“癌巢”,5例诊断为角、结膜乳头状瘤,2例诊断为角膜鳞状上皮细胞癌,与术后组织病理学诊断结果一致.结论 活体共聚焦显微镜检查对角膜上皮肿瘤术前诊断肿瘤性质提供重要依据.     

应用活体共聚焦显微镜观察糖尿病患者的角膜细胞

活体角膜共聚焦显微镜是一种无创、快速且全层面的技术,可以实时、动态地观察角膜组织的所有层面。共聚焦显微镜可以通过直接可视化检查角膜不同层面的形态和细胞密度。随着糖尿病患病率的不断上升,眼部并发症已经变得越发常见,并引起眼科临床工作者和科研工作者越来越多的兴趣和逐渐深入的研究。文章旨在通过采用活体角膜共聚焦显微镜观察糖尿病患者角膜组织各层的研究进展进行综述。     

In vivo confocal microscopy of pre-endothelial deposits

BACKGROUND: Deposits in various layers of the cornea might result from long-term medical therapy, photorefractive surgery, and longterm use of contact lenses or corneal dystrophies. METHODS: A 46-year-old woman was referred to our department with the suspected diagnosis of posterior polymorphous dystrophy. Slit-lamp biomicroscopy revealed bilateral small-sized deposits in the posterior part of the cornea. In vivo confocal microscopy was performed to evaluate these deposits in detail. RESULTS: In vivo confocal microscopy of the cornea identified hyperreflec-tive "dot-like" structures in the deep stromal layer and anterior to the endothelial cell layer. The morphology and number of keratocytes of the posterior stroma and of endothelial cells appeared normal. CONCLUSIONS: In vivo confocal microscopy is a very useful tool to analyze and visualize pre-endothelial deposits. Because there is no family history of corneal disease, the exact origin of the pre-endothelial deposits in our case remains unclear.     

应用活体共焦显微镜观察翼状胬肉组织结构

目的 应用活体共焦显微镜(德国海德堡视网膜激光断层扫描系统Ⅱ代与Rostock角膜模 块组成)对人眼翼状胬肉进行活体组织成像分析.方法 系列病例分析.采取2010年3月门诊就诊的翼 状胬肉患者中选择11例14只眼,应用活体共焦显微镜对翼状胬肉头部、颈部和体部进行检查,记录各部分图像,分析其组织形态特点.结果 应用共焦显微镜观察到翼状胬肉头部两种上皮结构:一种是翼状胬肉上皮细胞,包括浅表上皮细胞和深层上皮细胞.浅表上皮细胞呈边界高反光的不规则多边形,浅表上皮细胞间见核/浆比显著失调的不规则细胞,深层上皮细胞间可见大量成熟的朗格汉斯细胞浸润;另一种是角膜上皮细胞,细胞间散在炎性细胞浸润.头部角膜浅基质层可见一些基质细胞和层间走行的纤维组织.翼状胬肉颈部和体部的上皮细胞结构与头部胬肉上皮结构相似,颈部血管较正常结膜血管丰富,在头部形成栏栅样结构,未跨越透明角膜,同时可以看到血管中流动的血细胞.体部基质层清晰可辨,由大量纤维组织构成,走行与基质平面基本平行并相互交错成致密的网状结构,可见新生血管横跨其间.结论 活体共焦显微镜是一种非侵入性的组织观察方法,提供了翼状胬肉的活体、实时、动态的图像资料,是研究翼状胬肉组织结构的一种有效工具.

Abstract：

Objective To study the histology of the pterygia in vivo by confocal microscopy. Methods Series of case study. Fourteen pterygium eyes in 11 patients were selected for using in vivo confocal mieroscopy. Results Two kinds of epithelial cells were observed in the transitional zone of cornea and pterygium. Superficial epithelium cells of the pterygium characterized as polygon cells with brightly refractive border. There were many mature Langerhans cells and big irregular cells with significantly decreased nuclear/cytoplasm ratio within the Epithelial. Irregularly brightly reflective cells presented in the corneal epithelia. Fibrous tissue from the pterygium and cornea stroma cells were found in the superficial stroma. The blood vessels became dilated in the neck and body of pterygium, terminated at the head as palisade-like structures with lots of blood cells. The stroma of the pterygium body was filled with a lot of fibers which formed a dense net with neovascular across. Conclusions IVCM is a powerful technique for studying the vivo histology of the pterygium.Noninvasive technique for living tissue has provided in vivo, real-time, dynamic image data. It is a useful tool for investigating the structures ofpterygium.     

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.     

活体共焦显微镜对结膜滤过泡在细胞水平的研究

目的 应用活体共焦显微镜对小梁切除术后结膜滤过泡的组织病理学改变进行细胞水平的观察,研究结膜滤过泡形成的影响因素并进一步探讨滤过性手术的滤过机制.方法 对小梁切除术后3周至510周的59例患者共86只眼行裂隙灯、眼压及活体共焦显微镜检查.将受试眼的滤过泡分为两种类型:①功能性滤过泡型(43只眼);②非功能性滤过泡型(43只眼).根据术中是否使用过MMC再将各型滤过泡分为Normal组与MMC组.对各组滤过泡中微囊泡的大小、数量、结缔组织的密度进行统计学分析.结果功能性滤过泡中含有大量透明的微囊泡,上皮下存在比较疏松的结缔组织;而非功能性滤过泡中没有或仅有少许透明度较低的微囊泡,结缔组织极致密;功能性滤过泡中出现新生血管的比例远远小于非功能性滤过泡.与Normal组相比较术中联合应用MMC的功能性滤过泡中含有大量直径较大、壁薄的微囊泡,其上皮下结缔组织较疏松,各层组织内均可见大量高反光颗粒;而术中联合应用MMC的非功能性滤过泡仅结缔组织比Normal组疏松.结论 活体共焦显微镜检查能对结膜滤过泡进行诊断成像,是一种与来自离体组织学检查结果完全吻合的新的活体检查方法 ,对将来提高滤过性手术的成功率具有直接的指导作用.     

干燥综合征患者活体角膜组织学变化的研究

目的 应用共焦激光角膜显微镜观察干燥综合征(Sj(o)gren syndrome,SS)患者角膜上皮及上皮基底层下神经的组织学改变.方法 用共焦激光角膜显微镜对22例干燥综合征患者进行检查,同时选择年龄与性别与之相匹配的15名正常人作对照,并对两组人员行角膜敏感度检查.结果 (1)SS组与正常人对照组比较角膜敏感度下降(P=0.000). (2)两组间角膜上皮基底细胞密度的变化无统计学意义(P>0.05). (3)关于角膜卜皮基底层下神经纤维的改变:SS组神经数量比正常对照组增加(P=0.000);神经密度比正常对照组增加(P=0.046);神经弯曲度比正常对照组增加(P=0.001);神经分支数也比正常对照组增加(P=0.040). (4)神经弯曲度与角膜敏感度呈负相关(r=0.304,P=0.029).结论 SS患者的角膜上皮、上皮基底层下神经纤维以及角膜敏感度均发生了一系列改变.     

In vivo confocal microscopy of pre-Descemet's membrane corneal dystrophy

Pre-Descemet's membrane corneal dystrophy is clinically characterized by the presence of numerous tiny pleomorphic opacities located in the deep stroma immediately anterior to Descemet's membrane. A 35-year-old man, clinically diagnosed with pre-Descemet's corneal dystrophy, was examined by in vivo slit scan confocal microscopy. The pleomorphic structures containing dense hyperreflective inclusions in the posterior stroma were revealed in vivo. To the best of the authors' knowledge, it is consistent with the result of the previous histological study, but different from other reports using in vivo confocal microscopy.     

Assessing the sub-basal nerve plexus of the living healthy human cornea by in vivo confocal microscopy

The purpose of the study was to perform quantitative analysis of the sub-basal epithelial nerve plexus of healthy, living human cornea,using real time in vivo confocal microscopy and the analySIS software system. The study was based on in vivo confocalmicrostructural analysis of 50 eyes of 50 subjects, divided into two age groups: group 1 (n = 25)25 +/- 5 years of age, and group 2 (n = 25) 70 +/- 5 years of age. All subjects exhibited clinically healthy corneas. The overall nerve density was 632.35 +/- 287.57 microm/mm2 for group 1 and 582.39 +/- 327.13 microm/mm2 for group 2. The mean fibre dia-meter was measured at 0.52 +/- 0.23 microm for group 1 and at 0.56 +/- 0.27 microm for group 2. Beadings of the nerve fibres were recorded at a density of 213 +/- 123/mm for group 1 and 201 +/- 192/mm for group 2. Establishing standards for normal nerve density and morphology of the living human cornea at different ages may be beneficial, both in early detection and follow up of various corneal diseases and in post-surgical management following corneal surgery.     

不同光源共焦显微镜的活体正常角膜图像研究

目的 分别采用卤素灯光源共焦显微镜和激光光源共焦显微镜对中国正常成人活体角膜各层组织结构进行观察,对照研究其图像特征.方法 对35例(70只眼)中国成人(年龄18～55岁)中央部角膜分别采用卤素灯光源共焦显微镜和激光光源共焦显微镜进行检查,研究角膜各层结构的图像特点,并进行对比.结果 共焦显微镜检查的35例(70只眼)中,67只眼95.7％成功获得了角膜上皮翼状细胞层的图像,70只眼100％成功获得角膜上皮基底细胞层、前弹力层、前基质层、后基质层及角膜内皮细胞层的图像.结论 卤素灯光源共焦显微镜的放大倍数是1000倍,细胞放大倍数大,近似于初级电镜的放大倍数,但卤素灯光源的组织穿透力较弱,光源易衰减,所得的角膜各层细胞图像较激光角膜共焦显微镜的略微模糊,所得角膜各层图像总体颜色偏灰、结构偏模糊.激光角膜共焦显微镜的放大倍数是800倍,较卤素灯光源的共焦显微镜略小,但激光光源的组织穿透力强,所得角膜各层细胞图像较卤素灯光源的更清晰,而且在特定角度可获得非常类似角膜组织病理学切片的图像.     

激光共聚焦显微镜观察春季角结膜炎患者的角膜形态变化

目的 探讨应用活体激光共聚焦显微镜观察春季角结膜炎(VKC)患者的角膜形态变化.方法 观察型系列病例研究.选择2008年10月至2009年8月在复旦大学附属眼耳鼻喉科医院眼科就诊的26例双眼VKC患者,其中眼睑型13例、角膜缘型5例、混合型8例;另选择26名年龄、性别匹配的正常志愿者作为对照.使用活体激光共聚焦显微镜对患者右眼角膜进行检查,分别取中央角膜和上方周边部角膜为检查点,对所得图像进行记录和分析.利用内置细胞计数软件计算角膜各层组织的细胞密度,ImageJ软件分析神经密度、直径、分支数量及弯曲度.使用独立t检验比较VKC患者与正常人群角膜各层细胞密度的差异以及VKC患者周边与中央角膜细胞密度的差异;Fisher精确卡方检验比较VKC患者和对照组角膜上皮内郎格罕细胞浸润情况差异;方差分析比较VKC 3种亚型之间和不同病程之间各层细胞密度的差异;独立t检验和卡方检验分析VKC患者与正常人群之间的神经差异.结果 VKC患者的角膜形态学变化包括角膜上皮最表层的高亮多角形细胞缺失、上皮内和上皮下大量郎格罕细胞浸润及浅基质层内大量基质细胞活化.共焦显微镜下,角膜出现新生血管翳的患者表现为角膜上皮结膜化,基质内可见新生血管;合并圆锥角膜的患者,深基质内可见大量粗大的斜形或纵形暗纹.正常对照组的中央和周边部角膜上皮细胞密度分别为(6033.1±998.7)个/mm2和(6098.4±298.3)个/mm2,VKC患者分别为(5972.2±1148.2)个/mm2和(6178.5±318.9)个/mm2,差异无统计学意义(t=1.191,1.011;P=0.238,0.318);但VKC患者周边部上皮细胞密度显著高于中央部(t=2.249,P=0.03).正常对照组的中央和周边浅基质细胞密度分别(1001.4±125.3)个/mm2和(924.6±201.4)个/mm2,VKC患者分别为(1184.5±115.3)个/mm2和(1101.4±151.1)个/mm2,均显著高于正常对照(t=6.617,3.439;均P=0.001).正常对照与VKC患者中央角膜深基质层细胞密度分别为(537.7±42.6)个/mm2和(548.7±79.8)个/mm2,内皮细胞层细胞密度分别为(2985.7±401.2)个/mm2和(3021.5±383.3)个/mm2,两者比较均无明显差异(t=0.174,1.112;P=0.864,0.282).61.5%(16例)VKC患者的角膜上皮内发现郎格罕细胞浸润,显著高于正常人群(2例,7.7%)(x2=12.49,P=0.001).3种不同亚型VKC患者中,角膜缘型和混合型患者上皮内郎格罕细胞浸润情况较为严重.与正常对照组相比,VKC患者的上皮下神经密度和直径下降、弯曲度增加.结论 VKC主要累及角膜上皮层、上皮下神经及浅基质层.共焦显微镜对于VKC的分型诊断有一定辅助价值.

Abstract：

Objective To explore the morphological characteristics on cornea in patients with vernal keratoconjunctivitis(VKC)by the application of in vivo laser scanning confocal microscopy(LSCM).Methods The experimental design was retrospective observation case series(case control study).Twenty-six patients, each diagnosed as bilateral VKC, were enrolled in the study, among which 13 were tarsal form, 5 were bulbar form and the rest were mixed form. Nine patients had the clinical course less than one year, eight subjects longer than three years, and the rest between them. Another twenty-six healthy volunteers with matching age and gender were selected as normal control. All participants had their right eyes examined with the in vivo confocal microscopy ( HRT Ⅱ/RCM). Central cornea and superior peripheral cornea were chosen as the examination points. The images were recorded automatically and cellular density of each layer was analyzed by installed software. Software Image J was utilized to analyze the density, diameter, branch number and tortuosity of subbasal nerve fiber in VKC patients. Independent t test was performed to assess the differences on cellular density between VKC patients and normal control, as well as those between central and peripheral cornea in VKC patients. Fisher chi-square test was used to compare the infiltration rate of Langerhans cells in corneal epithelium between VKC patients and controls. ANOVA was applied to assess the differences in cellular density among three subtypes, as well as among different duration of VKC. Independent t-test and chi-square test were applied to analyze the parameters of subbasal nerve fiber. Results The morphological changes in cornea included the absence of superficial hyperreflective polygonal epithelial cells, infiltration of Langerhans cells in and(or) underneath corneal epithelium and activation of keratocytes in anterior stroma. Corneal epithelium conjunctivalization and stromal neovascularization could be identified in patients with corneal neovascular epithelium. Longitudinal or oblique dark striae could be found in the posterior stroma in patients with complicated keratoconus. The density of epithelial cells at central and peripheral cornea in healthy controls were (6033. 1 ± 998. 7) cells/mm2 and (6098. 4 ± 298. 3 ) cells/mm2, while that in VKC patients were (5972.2 ± 1148.2) cells/mm2 and (6178.5 ± 318.9) cells/mm2 respectively, the differences being no statistical significant between them (t = 1. 191 , 1. 011; P =0.238, 0. 318). However, it's found in VKC patients that cellular density at peripheral cornea was significantly higher than that at central area( t = 2. 249, P = 0. 03). The density of anterior stromal cells at central and peripheral cornea in healthy controls was (1001. 4 ± 125. 3) cells/mm2 and (924. 6 ± 201.4) cells/mm2, while that in VKC patients was (1184. 5 ± 115. 3 ) cells/mm2 and (1101.4 ± 151. 1) cells/mm2, the difference bearing no statistical significance(t =6. 617,3.439;P =0. 001). The density of posterior stromal cells in normal subjects and VKC patients was (537. 7 ± 42. 6) cells/mm2 and (548. 7 ± 79. 8) cells/mm2, that of endothelial cells was (2985. 7 ± 401. 2 ) cells/mm2 and (3021. 5 ± 383. 3) cells/mm2, respectively, neither difference had statistical significance (t = 0. 174, 1. 112; P = 0. 864, 0. 282 ) . Langerhans cell infiltration could be identified in 61.5% (16 cases) VKC patients, which was significantly higher than normal control (2 cases, 7. 7% ) (x2 = 12. 49, P = 0. 001 ). Furthermore, much intense Langerhans cells infiltration was found in bulbar form and mix form than tarsal form. (t = 6. 617, P = 0. 001). The density and diameter of subbasal nerve fiber in VKC patients decreased significantly than those in normal subjects, whereas the tortuosity increased significantly. Conclusions The morphological changes of cornea in VKC patients mainly involve corneal epithelium, subbasal nerve fiber and anterior stroma. In vivo LSCM is helpful in discriminating the subtypes of VKC.