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.     

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.     

Thiel-Behnke角膜营养不良活体共聚焦显微镜下特征及TGFBI基因突变分析

目的 研究Thiel-Behnke角膜营养不良患者临床表现、活体共聚焦显微镜下影像学特征及其TGFBI基因突变。 设计 前瞻性研究。 研究对象 Thiel-Behnke角膜营养不良家系35名成员。 方法 对该家系成员进行视力、裂隙灯显微镜、眼前节相干光断层扫描以及活体共聚焦显微镜检查；取外周静脉血制备血白细胞基因组DNA，PCR扩增TGFBI致病基因，分析患者基因序列变化情况。 主要指标 视力、裂隙灯显微镜检查、眼前节相干光断层扫描影像学观察、活体共聚焦显微镜检查，以及患者外周血基因序列突变分析。 结果 本家系4代35名家系成员中，10例患者表现为双眼角膜前弹力层及其附近可见弥漫性、灰白色混浊物沉积；病灶主要位于角膜中央区和角膜中周部的前弹力层，但浅基质层亦可见点状混浊病灶，呈线状或蜂窝状外观。眼前节相干光断层扫描显示患者角膜前弹力层及浅基质层出现连续、均匀一致的高反光沉积物，沉积物前缘（朝向上皮层）呈锯齿样；活体共聚焦显微镜在上皮层至浅基质层之间可见高反光、不规则、无定形物质沉积，病灶及周围组织未见炎性细胞浸润。外周血基因序列示该家系患者TGFBI基因第12外显子的第39密码子第2个碱基呈杂合子点突变G→A，导致精氨酸转变为谷氨酰胺取代（R555Q）。 结论 R555Q突变所致的Thiel-Behnke角膜营养不良活体共聚焦显微镜下影像学特征为角膜前弹力层高反光、不规则、蜂窝状、无定形物质沉积。     

共焦显微镜在角膜营养不良诊断中的应用

目的 探讨共焦显微镜在角膜营养不良诊断中的应用价值。设计 病例系列研究。研究对象 6例角膜营养不良，包括4例Reis-Bueckleas角膜营养不良、1例角膜斑点状营养不良、1例。Fuchs角膜内皮营养不良。方法 患者双眼行裂隙灯显微镜及共焦显微镜检查，选择病变在不同角膜层次的共焦显微镜图像，对角膜沉淀进行形态学评价，并与裂隙灯检查比较。主要指标角膜病变的裂隙灯显微镜及共焦显微镜图像。结果 共焦显微镜显示Reis-Bficklers角膜营养不良病变主要累及前部基质，包括角膜上皮、基底细胞及前弹力层；斑点状角膜营养不良病变仅累及基质层，而角膜上皮层及内皮层正常；在Fuchs角膜内皮营养不良中，可直接观察角膜小滴及角膜内皮情况。结论 共焦显微镜检查提供了一种评价角膜病变的方法，较裂隙灯显微镜的分辨率高。     

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。     

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。     

后部多形性角膜营养不良49例共聚焦显微镜影像学观察

目的:探讨49例后部多形性角膜营养不良(PPCD)患者在共聚焦显微镜(IVCM)下的影像学特征。方法:回顾性病例研究。收集我院2013-01/2021-01诊断为PPCD患者49例86眼,包括男32例,女17例,平均年龄42.5±22.9岁。所有患者均进行IVCM检查,分析角膜内皮细胞密度及不同类型病变的镜下特点。结果:所有患者病灶区内皮细胞数量均较外周细胞数量低。IVCM下病变呈1型囊泡型的有44眼(51%),表现为角膜内皮层的圆形或不规则弹坑样病灶,单个或多个出现;2型条带型16眼(19%),表现为边缘弧形凸起,部分可见似赘疣样影像散在或条索状分布;3型弥漫型26眼(30%),表现为内皮细胞大面积缺失,大部分区域内皮成像不清,呈匍匐样、条索样延展的凸起,成像较清晰处部分同条带型病灶。观察病例中有2例患者随访4～5a, IVCM下病灶均较前有变化,包括中央角膜内皮数量的减少和上皮层铁质沉着等。结论:IVCM能够显示PPCD患者各期内皮细胞和Descemet膜水平的特征性显微结构改变,通过观察其影像学特点可以为疾病提供有效的诊断价值。     

《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。     

活体共聚焦显微镜在微生物性角膜炎诊断中的应用进展

微生物性角膜炎(MK)是由细菌、病毒、真菌、棘阿米巴等微生物感染引起的角膜组织炎症病变,是导致角膜盲的重要原因之一。活体共聚焦显微镜(IVCM)是一种非侵入性成像技术,可通过IVCM快速、实时地获取角膜组织的高分辨率图像,在角膜疾病的诊断和临床研究中表现出独特的优势。近年来,随着学科交叉与融合,人工智能在辅助识别微生物性角膜炎IVCM图像的特征结构中崭露头角,对准确、快速地诊断MK具有重要的临床价值。因此,本文将从IVCM在诊断MK中的特征性表现以及人工智能在辅助医生诊断MK应用的进展两个方面进行综述,为进一步推动人工智能辅助共聚焦显微镜诊断角膜炎的诊疗做基础。     

《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。     

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.     

In vivo confocal microscopy findings in a patient with posterior amorphous corneal dystrophy

A 21-year-old man, with bilateral posterior amorphous corneal dystrophy, was studied by biomicroscopy, corneal topography and in vivo confocal microscopy. The best-corrected visual acuity was 6/21 in the right eye and 6/6.9 in the left eye. Biomicroscopy revealed bilateral, asymmetric, sheet-like opacification at the deep posterior stromal layer. The corneal topography displayed asymmetric against-the-rule astigmatism in the right eye and prominent steepening at the inferior paracentral cornea in both eyes. In vivo confocal microscopy of the corneas demonstrated microfolds and hyper-reflective layer at the posterior stroma just adjacent to the endothelial layer. The epithelium, Bowman's membrane, anterior stroma and the endothelial layer were normal. In vivo confocal microscopy is useful in evaluating the corneal dystrophies.     

Avellino角膜营养不良在共焦显微镜下的形态学特征

目的 探讨Avellino角膜营养不良（ACD）患者的共焦显微镜图像特点。方法对经基因型检测明确为ACD患者（R124H突变）的10例20眼角膜进行共焦显微镜检查，其中1例2眼为穿透角膜移植（PK）术后眼，一眼病变复发，另一眼无复发表现。并取颗粒状角膜营养不良I型（GCD I，R555W突变）患者9例18眼作为对照。结果ACD患者未手术眼共焦显微镜图像特点：基底上皮、Bowman膜及整个角膜基质均可见不同程度的异常反光改变；PK术后复发眼Bowman膜及紧邻其下的前基质可见由细小沉积物组成的不规则混浊；PK术后无复发表现眼角膜各层未见明显异常。GCD I型最显著的共焦显微镜图像特点为角膜基质内以大型沉积物为主。结论首次对经基因检测确诊的Avellino角膜营养不良患者的角膜进行活体共焦显微镜检查。ACD患者基底上皮、Bowman膜及整个角膜基质均可见不同程度的异常反光改变，但较GCD I型患者的沉积物为小。     

Differential diagnosis of corneal oedema assisted by in vivo confocal microscopy

The purpose of this study was to demonstrate microstructural differences between clinically similar, but aetiologically different, cases of corneal oedema in four subjects. In vivo confocal microscopy highlighted oedema of the basal epithelium, prominent nerve–keratocyte interactions, and typical ‘epithelialization’ of the endothelium in a case of iridocorneal endothelial syndrome; however, a similar microstructural appearance was observed in a case of presumed herpetic disciform keratitis. The latter diagnosis was subsequently revised on this basis. Confocal examination of Fuchs’ endothelial dystrophy demonstrated oedema of the basal epithelium, prominent wing cells, anterior stromal alterations, fibrosis of Descemet’s membrane and a typical ‘strawberry’ appearance of the endothelium. In contrast, in vivo microstructural examination of bilateral keratoconus with hydrops confirmed oedema mainly involving the epithelium and anterior stroma. In vivo confocal microscopy allows the clinician to observe the living cornea at a microstructural level and to better diagnose and differentiate borderline or unusual cases of corneal oedema.     

Imaging posterior polymorphous corneal dystrophy by in vivo confocal microscopy

To identify features of posterior polymorphous dystrophy (PPMD) by in vivo confocal microscopy, the corneas of a female patient with PPMD were examined using slit‐lamp biomicroscopy and slit‐scanning in vivo confocal microscopy. Characteristic endothelial vesicular and band lesions were seen clinically and easily identified using in vivo confocal microscopy. However, endothelial pleomorphism, an increased density and reflectance of posterior stromal keratocytes, and prominence of corneal nerves were also delineated. In vivo confocal microscopy enhances clinicopathological diagnosis and follow up of corneal dystrophies with subtle clinical presentations, such as PPMD.     

In vivo confocal microscopy in recurrent granular dystrophy in corneal graft after penetrating keratoplasty

Two case reports of recurrent granular dystrophy in corneal grafts after penetrating keratoplasty are presented. Slit-lamp examination and confocal microscopy (HRT II) were performed in two patients with recurrent granular dystrophy. All confocal microscopic findings of granular dystrophy were evaluated in the graft. Dystrophic lesions of the donor cornea presented the same confocal microscopic aspects in both eyes, and were similar to granular dystrophy lesions. Confocal microscopy is an imaging method that may provide new information on corneal microanatomy in dystrophies. It may be particularly useful in improving the early diagnosis of dystrophic lesions in corneal grafts.     

共焦显微镜在复发性角膜上皮糜烂综合征治疗中应用

目的：探讨共焦显微镜在指导长期配戴绷带式角膜接触镜治疗复发性角膜上皮糜烂综合征中的价值。

方法：随机选取2014-03/09就诊于我院36例36眼复发性角膜上皮糜烂综合征患者配戴绷带式角膜接触镜,观察戴镜前,戴镜后2、4、8、12wk眼部疼痛和刺激症状、角膜上皮愈合情况、共焦显微镜图像及不良反应情况。

结果：患者在配戴绷带式角膜接触镜后30min眼部的疼痛与刺激症状均得到不同程度缓解,完全缓解者26眼(72%),明显缓解者10眼(28%); 角膜上皮的平均愈合时间5.68±0.73d; 2、4、8、12wk时共焦显微镜下基底细胞形态、排列,上皮下神经纤维密度、走形日渐趋于正常; 临近角膜上皮基底细胞的前基质层炎细胞逐渐消退。观察期间未发现1例与配戴接触镜有关的并发症。

结论：共焦显微镜指导下绷带式角膜接触镜治疗复发性角膜上皮糜烂综合征可以避免过早摘镜,使基底膜与前弹力层紧密粘连,减少复发,是一种简便、安全、有效方法。     

Microstructural assessment of rare corneal dystrophies using real-time in vivo confocal microscopy

Purpose : To analyse and describe three cases of rare corneal dystrophy and highlight their in vivo microstructural features. Methods : Subject 1 was diagnosed with a posterior stromal fleck corneal dystrophy. Two of her three children were also affected. Subjects 2 and 3 exhibited an almost identical clinical appearance on biomicroscopic examination, such that both clinically were diagnosed as having pre‐Descemet’s dystrophies. All subjects underwent in vivo confocal microscopy and approximately 300 sequential digital images were obtained and analysed for each cornea. Results : In vivo confocal microscopy of subject 1 demonstrated an abnormal appearance of numerous large ovoid particles, measuring 50–70 μm in diameter in the mid and posterior stroma as well as smaller hyperreflective dot‐like intracellular deposits, of less than 1 μm diameter. Despite the near‐identical clinical appearance, subjects 2 and 3 could be clearly differentiated by in vivo confocal microscopy. Subject 2 exhibited small, irregular, optically dense particles, mainly in the anterior stroma, whereas subject 3 possessed classical involvement of the stroma immediately adjacent to Descemet’s membrane, with numerous regular, small, hyperreflective particles. Conclusions : The ability of in vivo confocal microscopy to localize and accurately measure various elements in different corneal layers may help to resolve whether abnormalities are intra‐ or extracellular, and aid clearer differentiation of rare corneal disorders.