Demonstration of bacterial cells and glycocalyx in biofilms on human tonsils
Demonstration of bacterial cells and glycocalyx in biofilms on human tonsils
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DOI：
10.1001/archotol.133.2.115
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发表时间：
2007-02-01
影响因子：
--
通讯作者：
Grote, Jan J.
中科院分区：
文献类型：
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作者：
Kania, Romain E.;Lamers, Gerda E. M.;Grote, Jan J.
目标：通过对扁桃体进行双重荧光染色的共聚焦激光扫描显微镜直接观察细菌和糖萼来展示人体组织中的粘膜生物膜，并将结果与扫描电子显微镜分析的结果进行比较。设计：前瞻性研究。设置：基于第三大学的转诊中心。患者：24个扁桃体来自慢性或复发性扁桃体炎的儿童。干预措施：扁桃体制备分析扫描电子显微镜和共聚焦激光扫描microscope.Main Outcome Measures：双荧光共聚焦激光扫描显微镜包括碘化丙啶染色检测细菌细胞和异硫氰酸荧光素伴刀豆球蛋白A染色检测糖萼基质。由3名研究者以盲法回顾性方式独立评价图像，分析图像的生物膜形态特征。需要所有观察员的共识，以证明存在的生物膜中的一个samples.Results：结果从分析使用扫描电子显微镜表明扁桃体上的生物膜形成的存在下，通过显示细菌细胞在微菌落。双染色技术，共聚焦激光扫描显微镜显示细菌细胞和糖萼基质，提供了视觉证据的生物膜上tonsills.Conclusion的存在：使用一种新的可视化方法在单切片的人类粘膜组织，生物膜的存在下，证明在大多数（17/24 [70.8%]）扁桃体炎患者的扁桃体。
objectives: To demonstrate mucosal biofilms in human tissue by direct visualization of bacteria and glycocalyx using confocal laser scanning microscopy with double fluorescent staining on tonsils and to compare the findings with the results of scanning electron microscopy analysis.Design: Prospective study.Setting: Tertiary university-based referral center.Patients: Twenty-four tonsils were obtained from children with chronic or recurrent tonsillitis.Interventions: Tonsils were prepared for analysis by scanning electronic microscopy and by confocal laser scanning microscopy.Main Outcome Measures: Double fluorescent staining for confocal laser scanning microscopy consisted of propidium iodide staining to detect bacterial cells and fluorescein isothiocyanate concanavalin A staining to detect the glycocalyx matrix. Images were analyzed for characteristic biofilm morphologic features by 3 investigators who evaluated the images independently in a blinded retrospective manner. Consensus of all observers was required to demonstrate the presence of a biofilm in a specimen.Results: Findings from analyses using scanning electronic microscopy suggested the presence of biofilm formations on tonsils by showing bacterial cells in micro-colonies. Double-staining technique using confocal laser scanning microscopy showed bacterial cells and the glycocalyx matrix, providing visual evidence for the presence of biofilms on tonsils.Conclusion: Using a novel visualization approach in single sections of human mucosal tissue, the presence of biofilms was demonstrated on tonsils in most (17/24 [70.8%]) patients with tonsillitis.