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目的观察铜绿假单胞菌实验室保存株与临床分离的菌株体外细菌生物膜的形成过程。方法通过FITC标记的刀豆球蛋白(FITC-ConA)和碘化丙啶(PI),结合激光共聚焦显微镜(confocal laser scanning microscopy,CLSM)摄取生物膜形成阶段不同层面的图片。结果培养3 d后可观测到生物膜中的多糖开始形成,培养第7 d,多糖基质逐渐增多,细菌趋向化聚集,膜结构致密,其内部存在相互交通的间隙及孔道;CLSM结合FITC-ConA和PI标记,实现了对铜绿假单胞菌菌株生物膜动态形成过程的观察。结论铜绿假单胞菌生物膜可在体外支持物上形成并成熟;临床新分离株较实验室保存株更易形成生物膜。
Objective To investigate the in vitro bacterial biofilm formation of Pseudomonas aeruginosa laboratory-preserved strains and clinical isolates. Methods FITC-ConA and PI were combined with confocal laser scanning microscopy (CLSM) to capture the different layers of the biofilm formation phase. Results After cultured for 3 days, the polysaccharide in biofilm began to form. On the 7th day after culture, the polysaccharide matrix gradually increased, the bacteria tended to aggregate, the membrane structure was dense, and there were gaps and channels in the internal communication. CLSM combined with FITC-ConA And PI markers to achieve the dynamic observation of biofilm formation of Pseudomonas aeruginosa strains. Conclusion Pseudomonas aeruginosa biofilms can be formed and matured on in vitro supports. New clinical isolates are more biofilm-forming than laboratory-preserved strains.