摘要
目的 建立变异链球菌生物膜模型 ,用激光共聚焦扫描显微镜 (CLSM)观察变异链球菌生物膜结构。方法 在盖玻片上分别形成 6、12、18、2 4、4 8、72h变异链球菌生物膜 ,将得到的各时段生物膜荧光染色后 ,用CLSM观察生物膜的断层扫描图像、生物膜厚度、每层红光绿光的面积 ,计算生物膜中细菌密度和活菌百分比 ,用软件处理扫描数据 ,得到生物膜的三维重建图像。结果 变异链球菌生物膜具有空间立体结构 ,形态多样 ,其中细菌密集 ,由死细菌和活细菌组成 ,还有丰富的基质和管道系统。 2 4h生物膜平均厚度最大 ,生物膜内层、中间层的细菌密度相对较大 ,而外层较低 ,72h内各时间段生物膜中活菌百分比由内往外逐渐增加。结论 变异链球菌生物膜有一定的厚度 ,具有三维立体空间结构 ,结构形态具有多样、不均质、开放的特点。
Objective To observe the architecture of Streptococcus mutans (S.mutans) biofilms utilizing of an experimental model of S.mutans biofilms in confocal laser scanning microscopy (CLSM). Methods S.mutans biofilms were formed on glass slices in 6, 12, 18, 24, 48, 72 h respectively. The scanning picture, depth and the area of green and red light per layer of biofilms were observed by CLSM. Bacterial density and the percentage of vital bacteria were calculated. The three-dimensional reconstruction of biofilm pictures were obtained by special software of scanning data of biofilms. Results It was shown that S.mutans biofilm had three-dimensional spatial architecture with various forms. A lot of vital and dead bacteria gathered densely in biofilms. Bacteria were embeded in rich extracellular polymers. There were abundant channels in biofilms. Among all the biofilms at every time slot we observed, those of 24 h were the thickest in terms of average level. Bacterial density was higher at both the interior and middle layers than at the outer layer. As for the biofilm within 72 h, the percentage of vital bacteria at every time slot increased from the inner layer to the outer. Conclusion S.mutans biofilm had a certain degree of thickness. Its structure was three-dimensioned, heterogeneous, openness and of various forms.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第2期103-106,共4页
Chinese Journal of Microbiology and Immunology
基金
教育部留学回国人员科研启动基金资助项目( 0 1GJHG0 2 )
