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.     

中国人正常角膜的共焦显微镜检查

摘要目的探讨正常人共焦显微镜下角膜各层细胞的活体细胞形态学特征及密度.方法对28例(年龄36.6士14.5岁,范围19～68岁)41眼(男14眼,女27眼)正常人中央部角膜进行活体共焦显微镜检查,描述角膜各层结构的图象特点,并分析角膜各层细胞的密度与年龄的相关性.结果上皮表层细胞排列疏松,边界清楚,核反光较强;基底层细胞排列紧密,胞质和核反光弱,偶尔可见点状细胞核,细胞平均密度为5724.41士562.5个/mm2.Bowman's膜为无一定形状的膜状物,仅可分辨的结构是上皮下的念珠状神经纤维.基质层中可见相对较暗的背景下明亮的角膜细胞核.前基质内有放射状分布的神经纤维.前基质层角膜细胞平均密度为1099.10士164.90个/mm     

佩戴角膜接触镜后角膜变化的激光共焦显微镜观察

目的 应用激光共焦显微镜对长期佩戴角膜软性接触镜的患者活体角膜的组织结构变化进行观察.方法 用激光共焦显微镜对长期佩戴角膜软性接触镜的15例患者进行检查,并选择未戴角膜接触镜者11例进行对照,对两组结果进行比较.结果 1.佩戴角膜软性接触镜组与对照组相比,基底上皮细胞密度减少,为3705.00±447.62个/mm2(P＜0.05),角膜上皮层厚度变薄,为54.3±8.44μm(P＜0.05),并有部分剥脱.2.佩戴角膜软性接触镜组角膜内出现白色点状物,朗汉氏细胞数目多,成树枝状改变.3.角膜软性接触镜组与对照组相比神经纤维数量及密度无明显变化(P＞0.05),但曲折度增加,有分支出现(P＜0.05).结论 佩戴角膜软性接触镜,角膜组织可发生一系列改变,激光共焦显微镜与传统光学共焦显微镜相比,图象清晰,深度定位准确,在疾病的早期诊断、治疗和研究中将起重要作用.     

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。     

人体角膜内皮细胞的共焦显微镜研究

目的应用共焦显微镜观察活体角膜内皮细胞的形态学特征。方法应用共焦显微镜观察正常人（20例）、角膜炎（12例）、角膜移植术后（19例）、葡萄膜炎（5例）、大疱性角膜病变（3例）、角膜白斑（6例）、角膜营养不良（7例）、青光眼（10例）、圆锥角膜（17例）等患者的角膜内皮,分析所得活体角膜内皮图象,总结其形态学特征。结果活体共焦显微镜下角膜内皮的图象主要有8种：①完全正常角膜内皮。②水肿角膜内皮。③失代偿角膜内皮。④“激活”角膜内皮。⑤排斥角膜内皮。⑥有KP角膜内皮。⑦有“疣状物”的角膜内皮。⑧“双核”角膜内皮细胞。结论共焦显微镜能对活体角膜内皮进行实时、无创、准确以及可重复、可比较的观察。     

共焦显微镜在角膜病的应用

共焦显微镜（ｃｏｎｆｏｃａｌｍｉｃｒｏｓｃｏｐｙ）是一种在四维（Ｘ，Ｙ，Ｚ轴和实时）观察活体角膜细胞水平，且无创伤的显微镜，其特点是无论在透明或混浊的角膜均能在三维空间和实时条件下显示角膜细胞结构和病理改变，尤其对角膜棘阿米巴，病毒感染等能在病因学上的作出诊断，对角膜移植术后的排斥反应能早期发现，对角膜的各种创伤愈合亦能观察到细胞，神经的动态改变，共焦显微镜被认为是继角膜内皮镜后，更具意义的新型角     

角膜内皮炎共焦显微镜形态学分析

目的:观察角膜内皮炎的共焦显微镜下形态学特征。方法:应用Confoscan 4.0共焦显微镜对24例24眼角膜内皮炎患者的角膜进行扫描检查,记录并分析各层角膜图像。结果:所有患者前部基质混浊,角膜深基质层可见基质细胞排列紊乱及条索状高反光结构,深基质层中还可见低反光带为后弹力层皱褶,角膜内皮细胞前可见斑片状大小不等的高反光结构,病变区角膜内皮细胞水肿、变性呈无结构暗区,内皮细胞呈多形性改变。4眼角膜上皮细胞边界不清,排列疏松,细胞较大,细胞核呈高反光结构,其中可见泡状暗区。结论:共焦显微镜可活体检查角膜内皮炎患者角膜组织各层结构,起到类似病理组织切片的作用;角膜内皮炎以深基质层及内皮细胞层损害为特征;共焦显微镜检查对角膜内皮炎具有一定的参考价值。     

共焦显微镜在角膜疾病中的临床应用

本文对一种新型的光学显微镜－共焦显微镜,在角膜病中的临床指导意义做了详细的报道。主要介绍了共焦显微镜的原理,使用方法,结构,并总结了其对正常人体角膜细胞研究的结果,概述了其对棘阿米巴角膜炎,角膜营养不良,角膜激光切削术,角膜移植术后等角膜疾病的临床研究结果,发现共焦是微镜是独特的,先进的,分辨更好的细胞水平研究的新型仪器、对角膜疾病的早期诊断,早期治疗有重要的临床意义。     

共焦激光角膜显微镜在真菌性角膜炎诊断中的应用

目的评价共焦激光角膜显微镜在临床诊断真菌性角膜炎中的应用价值。方法用共焦激光角膜显微镜对临床拟诊为真菌性角膜炎的26例患者进行检查,同时行涂片检查真菌菌丝,对两组结果进行比较。结果26例患者中,根据病史、临床表现24例确诊为真菌性角膜炎,其中共焦激光角膜显微镜的确诊率为95.83%(23/24),角膜刮片的确诊率为83.33%(20/24)。结论共焦激光角膜显微镜是一种快速、有效、无损伤的活体检查方法,在真菌性角膜炎的早期诊断、治疗和研究中将起重要作用。     

暗场显微镜联合角膜激光共焦显微镜在非典型真菌性角膜溃疡的应用研究

目的 对非典型的真菌性角膜溃疡患者(角膜刮片取材,涂片镜检不能确诊的真菌性角膜溃疡),采用暗场显微镜联合角膜激光共焦显微镜检查进行观察研究,并归纳总结其临床特点.方法 23例(23只眼)非典型的真菌性角膜溃疡患者,在角膜刮片取材培养前采用激光角膜共焦显微镜对溃疡灶进行活体检查,然后进行角膜刮片取材,涂片常规镜检和暗场显微镜检查,研究其图象特点,并进行对比研究.结果 非典型真菌性角膜溃疡灶中的真菌菌丝在活体角膜激光共焦显微镜中结构不完整、成像不清晰;在常规镜检下,成像不清甚至难以成像;但采用暗场显微镜检查能发现清晰的真菌菌丝.结论 暗场显微镜观察非典型真菌性角膜溃疡有着独特的优点,该方法联合常规涂片镜检及共焦显微镜检查能提高对非典型真菌性角膜溃疡的诊断水平.     

应用共聚焦显微镜观察正常老年人角膜形态学的特点

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。     

In vivo confocal microscopy of human cornea covered with human amniotic membrane

Purpose  Amniotic membrane transplantation has been widely performed to reconstruct the surface of the eye and treat chemical burns or epithelial defects. However, we have difficulty observing the cornea through the opaque transplanted amniotic membrane by slit-lamp biomicroscopy. We investigated the use of confocal microscopy for observation of human corneas covered with amniotic membrane. Methods  Human amniotic membrane was placed onto the normal corneas of five volunteers aged 22–24 years. Then, all layers of the covered corneas were observed by in vivo confocal microscopy. Results  Confocal microscopy displayed the epithelium, basement membrane, and stroma of the amniotic membrane. It also displayed the corneal epithelium. Furthermore, corneal stromal keratocytes and the corneal endothelium were clearly observed through the amniotic membrane by confocal microscopy. Conclusions  We demonstrated that in vivo confocal microscopy enabled us to observe all layers of corneas covered with amniotic membrane in normal human eyes. Our findings suggest that confocal microscopy may have advantages for clinical examination of the ocular surface, including all layers of the cornea.     

In vivo corneal confocal microscopy in keratoconus

PURPOSE: To evaluate the corneas of keratoconic subjects using in vivo confocal microscopy. METHODS: Slit scanning confocal microscopy was used to evaluate the central cornea of one eye of each of 29 keratoconic subjects (mean age 31 +/- 10 years; range 16-49 years). Quantitative aspects of corneal morphology were compared against data from control subjects. RESULTS: Compared with normal control corneas, epithelial wing cell nuclei were larger (p < 0.0001) and epithelial basal cell diameter was larger (p < 0.05) in the keratoconic cornea. Many of the keratoconic corneas investigated showed increased levels of stromal haze and reflectivity, which appeared to be related to the presence of apical scarring on slit lamp examination. A grading scale was devised to quantify the levels of haze. This scale was shown to provide a measure of the level of scarring present. The anterior keratocyte density (AKD) and posterior keratocyte density were 19% lower (p < 0.0001) and 10% lower (p = 0.004) than in controls, respectively. The reduction in AKD was significantly associated with three factors: a history of atopy, eye rubbing and the presence of corneal staining. The mean endothelial cell density in keratoconus was 6% greater than that of normal controls (p = 0.05). The level of endothelial polymegethism was shown not to be different between keratoconic subjects and matched controls (paired t-test: t = 1.82, p = 0.08). CONCLUSIONS: Confocal microscopy demonstrates significant quantitative alterations of corneal morphology in keratoconus.     

翼状胬肉在激光共焦显微镜下的观察研究

目的:应用激光共焦显微镜HRT3-RCM对翼状胬肉进行细胞学观察。方法:对20例20眼翼状胬肉的患者用HRT3-RCM进行观察。将其与对侧正常眼(对照组)进行比较。结果:翼状胬肉区域往往见不到Vogt栅栏结构,取而代之的是大量的纤维组织和丰富血管。结论:HRT3-RCM是利用激光作为光源的新一代共焦显微镜,是一种快速、有效、无创的检查方法,可用于翼状胬肉组织的细胞学观察。     

In vivo confocal laser-scanning microscopy to characterize wound repair in rabbit corneas after collagen cross-linking

Background: Collagen cross‐linking using the photosensitizer riboflavin combined with ultraviolet A light was developed to stiffen the cornea by increasing its mechanical and biochemical stability. Investigation of post‐treatment events, such as wound healing, is important to evaluate possible risks and to optimize treatment protocols. This in vivo confocal laser‐scanning microscopy study in rabbits was conducted to provide a quantitative and qualitative analysis of corneal wound repair over 16 weeks following collagen cross‐linking. Methods: Six New Zealand White rabbits underwent riboflavin/ultraviolet A cross‐linking. In vivo confocal laser‐scanning microscopy using a Heidelberg Retina Tomograph equipped with a Rostock Cornea Module was performed preoperatively and at 2, 4, 8, 12 and 16 weeks postoperatively. Results: From 2 weeks onwards the epithelium demonstrated no abnormalities. Evidence of inflammation was visualized in the intermediate, basal cells and Bowman's membrane. Nerve fibre regeneration was first noted at 12 weeks. Keratocyte activation and hyperreflective extracellular matrix were observed consistently, but by 16 weeks keratocyte activation was diminished, and extracellular matrix resumed normal reflectivity. Cell density in the posterior stroma and endothelium regained preoperative values by 4 weeks, although anterior stroma keratocyte cell density was still reduced by about 10% at 16 weeks. Conclusions: Complete qualitative and quantitative characterization of corneal wound repair was achieved by in vivo confocal laser‐scanning microscopy over 16 weeks following collagen cross‐linking in rabbits. In terms of assessing the ever‐increasing range of cross‐linking protocols, in vivo confocal laser‐scanning microscopy may contribute to minimizing the number of experimental animals, because multiple examinations of the same cases are possible over time.