离体及活体念珠菌在共聚焦显微镜下的特征

目的：应用活体激光共聚焦显微镜（IVCM）观察离体培养及活体状态下念珠菌孢子及假菌丝的典型 特征，为念珠菌性角膜炎的早期诊断提供依据。方法：比对观察研究。收集2016年1月到2019年12月 在山东第一医科大学附属眼科医院被诊断为念珠菌性角膜炎的26例（26眼）患者资料，同时取白色 念珠菌和近平滑念珠菌进行实验室真菌培养，应用IVCM分别观察离体及活体状态下念珠菌孢子及 假菌丝的典型形态。结果：离体与活体状态下念珠菌孢子均呈排列均匀的点状结构，形状类圆形， 直径约2～5 μm，边界清晰；假菌丝呈高反光线状结构。活体状态下孢子反光明显增强，18眼呈分散 排列点状强反光结构，11眼呈团雾状中强反光结构；假菌丝形态变异较大，2眼菌丝形体细长，5眼 呈芽孢结构，可见分支，1眼呈高对比度的不规则细长短棒状，1眼可同时观察到不规则细长短棒状 和芽孢结构。白色念珠菌假菌丝结构较细长，长度约20～150 μm，直径约2～10 μm，与母细胞连接 处收缩呈缢痕；近平滑念珠菌假菌丝短粗，长度约30～100 μm，直径2～5 μm，部分可见高亮的分支 芽孢结构。结论：念珠菌性角膜炎在IVCM下呈现典型的孢子和假菌丝结构，离体和活体状态下孢 子形态较为一致，而假菌丝形态变异较大，IVCM可用于念珠菌性角膜炎的早期诊断。

A Study of the Characteristics of Candida with In Vitro and In Vivo Confocal Microscopy

Objective: To use in vivo laser confocal microscopy (IVCM) to observe the in vitro culture and living characteristics of candida spores and pseudohyphaes, and to provide abasis for the early diagnosis of candida keratitis. Methods: The data of 26 patients (26 eyes) who were diagnosed with candida keratitis in the Eye Hospital of Shandong First Medical University from January 2016 to December 2019 wascollected. At the same time, candida albicans and candida parapsilosis were taken for laboratory fungal culture, and the typical morphology of candida spores and pseudohyphae in vitro and in vivo were observed by IVCM. Results: Candidaspores in vitro and in vivo presented uniformly in the shape of round spots with a diameter of 2-5 μm and a clear boundary. Pseudohyphae have a highly reflective linear structure. The reflection of spores was significantly enhanced in vivo. Eighteen eyes showed a scattered arrangement of spot-like, strongly reflective structure, and 11 eyes showed a dense fog structure. The morphology of pseudomycelia varied greatly. The mycelia in 2 eyes were slender, the spore structure was present in 5 eyes, and branches were visible. In 1 eye, an irregular thin and long rod shape with high contrast was observed, and in another eye, an irregular thin and long rod shape and spore structure could be observed simultaneously. Candida albicans pseudohyphae were relatively slender, with a length of 20-150 μm and a diameter of 2-10 μm. Pseudohyphae of candida parapsilosis were short and thick, with a length of about 30-100 μm and a diameter of 2-5 μm. Part of the branch spore structure was highlighted. Conclusions: Candida keratitis showstypical spore and pseudohyphae structures with IVCM. The morphology of candida keratitis spores in vitro and in vivo are consistent with observations under IVCM, but the morphology of pseudohyphaes are relatively different. IVCM can be used for the early diagnosis of candida keratitis.